Serine protease inhibitors

ABSTRACT

Compounds of formula (I)  
                 
 
     in which R 2 , X, Y, Cy, L and Lp(D) n  have the meanings given in the specification, are inhibitors of the serine protease, Factor Xa and are useful in the treatment of cardiovascular disorders.

[0001] This invention relates to compounds which are inhibitors ofserine proteases and to pharmaceutical compositions thereof and theiruse in the treatment of the human or animal body.

[0002] The serine proteases are a group of proteolytic enzymes whichhave a common catalytic mechanism characterized by a particularlyreactive Ser residue. Examples of serine proteases include trypsin,tryptase, chymotrypsin, elastase, thrombin, plasmin, kallikrein,Complement C1, acrosomal protease, lysosomal protease, cocoonase,α-lytic protease, protease A, protease B, serine carboxypeptidase II,subtilisin, urokinase, Factor VIIa, Factor IXa, and Factor Xa. Theserine proteases have been investigated extensively over a period ofseveral decades and the therapeutic value of inhibitors of serineproteases is well understood.

[0003] Serine protease inhibitors play a central role in the regulationof a wide variety of physiological process including coagulation,fibrinolysis, fertilization, development, malignancy, neuromuscularpatterning and inflammation. It is well known that these compoundsinhibit a variety of circulating proteases as well as proteases that areactivated or released in tissue. It is also becoming clear that serineprotease inhibitors inhibit critical cellular processes, such asadhesion, migration, free radical production and apoptosis. In addition,animal experiments indicate that intravenously administered serineprotease inhibitors, variants or cells expressing serine proteaseinhibitors, provide a protective effect against tissue damage.

[0004] Serine protease inhibitors have also been predicted to havepotential beneficial uses in the treatment of disease in a wide varietyof clinical areas such as oncology, neurology, haematology, pulmonarymedicine, immunology, inflammation and infectious disease.

[0005] In particular serine protease inhibitors may be beneficial in thetreatment of thrombotic diseases, asthma, emphysema, cirrhosis,arthritis, carcinoma, melanoma, restenosis, atheroma, trauma, shock andreperfusion injury.

[0006] Thus for example an inhibitor of Factor Xa has value as atherapeutic agent as an anticoagulant, e.g. in the treatment andprevention of thrombotic disorders. The use of a Factor Xa inhibitor asan anticoagulant is desirable in view of the selectivity of its effect.Many clinically approved anticoagulants have been associated withadverse events owing to the non-specific nature of their effects on thecoagulation cascade.

[0007] Also, there are well-known associations of α1 protease inhibitordeficiency with emphysema and cirrhosis and C1 esterase inhibitordeficiency with angioedema.

[0008] It has now been found that certain aromatic compounds carryingbulky lipophilic side chains are particularly effective as inhibitors ofserine proteases, especially proteases with negatively charged P1specificity pockets, and most especially the serine proteases thrombin,and most importantly Factor Xa. The Factor Xa inhibitors of thisinvention are potentially useful for the prophylaxis or treatment ofthrombotic disorders such as amongst others venous thrombosis, pulmonaryembolism, arterial thrombosis, myocardial ischaemia, myocardialinfarction, and cerebral thrombosis. They potentially have benefit inthe treatment of acute vessel closure associated with thrombolytictherapy and restenosis, e.g. after transluminal coronary angioplasty orbypass grafting of the coronary or peripheral arteries and in themaintenance of vascular access patency in long term hemodialysispatients.

[0009] Factor Xa inhibitors of this invention may, with benefit, formpart of a combination therapy with an anticoagulant with a differentmode of action or with a thrombolytic agent.

[0010] It has been reported in WO99/11658 and WO99/11657 that certainbenzamidine and aminoisoquinoline derivatives carrying a bulkylipophilic side chain are excellent inhibitors of serine proteases.Unfortunately, it has since been found that benzamidine compounds of WO99/11658 in general demonstrate poor oral bioavailability.

[0011] Surprisingly, it has now been found that certain other aromaticcompounds also show inhibitory activity against serine proteases, inparticular Factor Xa, despite the lack of the amidino or1-aminoisoquinoline functionality previously believed to be crucial foractivity as a factor Xa inhibitor. Many of these compounds also possessother structural features that further distinguish them from thecompounds of WO99/11658 and WO99/11657.

[0012] Where compounds of the invention have been tested, they havegenerally demonstrated superior oral bioavailability in comparison withbenzamidines disclosed in WO 99/11658. Also, it has been found that thecompounds of the invention perform excellently in the prothrombin timeassay (PT) when compared to aminoisoquinolines of similar factor Xaactivity and structure. The PT assay is a coagulation assay and it iswidely accepted that direct acting Factor Xa inhibitors which performwell in the PT assay are more likely to be good antithrombotics.

[0013] In WO99/09053 certain 2-aminobenzamide compounds are disclosed aspotential motilin receptor antagonists and in U.S. Pat. No. 3,268,513similar 2-aminobenzamide compounds are suggested as potentialantibacterial agents. However, the novel compounds of the presentinvention have not before been suggested as potential serine proteaseinhibitors.

[0014] Thus viewed from one aspect the invention provides a serineprotease inhibitor compound of formula (I)

[0015] wherein:

[0016] R₂ is a 5 or 6 membered aromatic carbon ring optionallyinterrupted by a nitrogen, oxygen or sulphur ring atom, optionally beingsubstituted in the 3 and/or 4 position (in relation to the point ofattachment of X—X) by halo, nitro, thiol, haloalkoxy, hydrazido,alkylhydrazido, amino, cyano, haloalkyl, alkylthio, alkenyl, alkynyl,acylamino, tri or difluoromethoxy, carboxy, acyloxy, MeSO₂— or R₁, orthe substituents at the 3 and 4 positions taken together form a fusedring which is a 5 or 6 membered carbocyclic or heterocyclic ringoptionally substituted by halo, haloalkoxy, haloalkyl, cyano, nitro,amino, hydrazido, alkylthio, alkenyl, alkynyl or R_(1j), and optionallysubstituted in the position alpha to the X—X group (i.e. 6 position fora six membered aromatic ring etc) by amino, hydroxy, halo, alkyl,carboxy, alkoxycarbonyl, cyano, amido, aminoalkyl, alkoxy or alkylthiowith the proviso that R₂ cannot be aminoisoquinolyl;

[0017] each X independently is a C, N, O or S atom or a CO, CR_(1a),C(R_(1a))₂ or NR_(1a) group, at least one X being C, CO, CR_(1a) orC(R_(1a))₂;

[0018] each R_(1a) independently represents hydrogen or hydroxyl,alkoxy, alkyl, aminoalkyl, hydroxyalkyl alkoxyalkyl, alkoxycarbonyl,alkylaminocarbonyl, alkoxycarbonylamino, acyloxymethoxycarbonyl oralkylamino optionally substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl;

[0019] R₁ is as defined for R_(1a), provided that R₁ is notunsubstituted aminoalkyl;

[0020] Y (the α-atom) is a nitrogen atom or a CR_(1b) group;

[0021] Cy is a saturated or unsaturated, mono or poly cyclic, homo orheterocyclic group, preferably containing 5 to 10 ring atoms andoptionally substituted by groups R_(3a) or R_(3i)X_(i);

[0022] each R_(3a) independently is R_(1c), amino, halo, cyano, nitro,thiol, alkylthio, alkylsulphonyl, alkylsulphenyl, triazolyl, imidazolyl,tetrazolyl, hydrazido, alkylimidazolyl, thiazolyl, alkylthiazolyl,alkyloxazolyl, oxazolyl, alkylsulphonamido, alkylaminosulphonyl,aminosulphonyl, haloalkoxy, haloalkyl, a group of the formula—C(X³)N(R¹¹)R¹² (wherein X³ is O or S; and R¹¹ and R¹² are independentlyselected from hydrogen, methyl or ethyl or together with the nitrogenatom to which they are attached form a pyrrolidin-1-yl, piperidin-1-ylor morpholino group), or —OCH₂O— which is bonded to two adjacent ringatoms in Cy;

[0023] X_(i) is a bond, O, NH or CH₂;

[0024] R_(3i) is phenyl, pyridyl or pyrimidinyl optionally substitutedby R_(3a);

[0025] R_(1b), R_(1c) and R_(1j) are as defined for R_(1a); and

[0026] -L-Lp(D)_(n) is of the formula:

[0027] in which R_(r) is —(CH₂)_(c)—R_(c), —CHR_(e)R_(f),—CH₂—CHR_(e)R_(f), —CH₂—CH₂—CHR_(e)R_(f), or R_(g) in which c is 1 or 2;R_(c) is thienyl, thiazolyl (which may bear an amino substituent),isothiazolyl, oxazolyl, isoxazolyl, pyrazolyl, imidazolyl, pyridyl(which may bear an alkylsulphonyl, aminosulphonyl, alkylaminosulphonyl,alkylaminocarbonyl, amino, amido, (1-4C)alkoxycarbonyl, carboxy,acetylamino, chloro, fluoro, cyano, (1-3C)alkyl, trifluoromethyl,methoxy, ethoxy, nitro, hydroxy, alkylsulphonylamino, triazolyl ortetrazolyl substituent), pyrimidinyl, pyridazinyl, pyrazinyl or phenyl(which may bear a methyl, methylamino, dimethylamino, carboxy,dialkylaminosulphonyl, alkylsulphonyl, aminosulphonyl,alkylaminosulphonyl, alkylaminocarbonyl, amino, amido, alkoxycarbonyl,acetylamino, chloro, fluoro, cyano, methoxy, ethoxy, nitro, hydroxy,alkylsulphonylamino, triazolyl or tetrazolyl substituent); each of R_(e)and R_(f) independently is hydrogen or C₁₋₃alkyl; or CHR_(e)R_(f) iscyclopentyl (which may bear a hydroxy, amino, (1-3C)alkoxy,(1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy, methoxycarbonyl orethoxycarbonyl substituent at the 3- or 4-position), cyclohexyl (whichmay bear a hydroxy, amino, (1-3C)alkoxy, (1-3C)hydroxyalkyl,(1-3C)alkyl, carboxy, methoxycarbonyl or ethoxycarbonyl substituent atthe 3- or 4-position), tetrahydropyran-4-yl, tetrahydrothiopyran-4-yl,pyrrolidin-3-yl (which may bear a hydroxy, amino, (1-3C)alkoxy,(1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy, methoxycarbonyl orethoxycarbonyl substituent at the 1-position), piperidin-4-yl (which maybear a hydroxy, amino, (1-3C)alkoxy, (1-3C)hydroxyalkyl, (1-3C)alkyl,carboxy, methoxycarbonyl or ethoxycarbonyl substituent at the1-position), or indan-2-yl; and R_(g) is 2-methylsulphonylphenyl whichmay bear a 4-fluoro substituent or R_(g) isλ⁶-1,1-dioxobenzo[b]thiophen-7-yl;

[0028] or a physiologically-tolerable salt thereof (e.g. a halide,phosphate or sulfate salt or a salt with ammonium or an organic aminesuch as ethylamine or meglumine); provided that Lp(D)n is not of theformula (K):

[0029] wherein X₂ is fluoro or hydrogen.

[0030] In another aspect the invention relates to a serine proteaseinhibitor compound of formula (I)

[0031] wherein:

[0032] R₂ is a 5 or 6 membered aromatic carbon ring optionallyinterrupted by a nitrogen, oxygen or sulphur ring atom, optionally beingsubstituted in the 3 and/or 4 position (in relation to the point ofattachment of X—X) by halo, nitro, thiol, haloalkoxy, hydrazido,alkylhydrazido, amino, cyano, haloalkyl, alkylthio, alkenyl, alkynyl,acylamino, tri or difluoromethoxy, carboxy, acyloxy, MeSO₂— or R₁, orthe substituents at the 3 and 4 positions taken together form a fusedring which is a 5 or 6 membered carbocyclic or heterocyclic ringoptionally substituted by halo, haloalkoxy, haloalkyl, cyano, nitro,amino, hydrazido, alkylthio, alkenyl, alkynyl or R_(1j), and optionallysubstituted in the position alpha to the X—X group (i.e. 6 position fora six membered aromatic ring etc) by amino, hydroxy, halo, alkyl,carboxy, alkoxycarbonyl, cyano, amido, aminoalkyl, alkoxy or alkylthiowith the proviso that R₂ cannot be aminoisoquinolyl;

[0033] each X independently is a C, N, O or S atom or a CO, CR_(1a),C(R_(1a))₂ or NR_(1a) group, at least one X being C, CO, CR_(1a) orC(R_(1a))₂;

[0034] each R_(1a) independently represents hydrogen or hydroxyl,alkoxy, alkyl, aminoalkyl, hydroxyalkyl alkoxyalkyl, alkoxycarbonyl,alkylaminocarbonyl, alkoxycarbonylamino, acyloxymethoxycarbonyl oralkylamino optionally substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl;

[0035] R₁ is as defined for R_(1a), provided that R₁ is notunsubstituted aminoalkyl;

[0036] Y (the α-atom) is a nitrogen atom or a CR_(1b) group;

[0037] Cy is a saturated or unsaturated, mono or poly cyclic, homo orheterocyclic group, preferably containing 5 to 10 ring atoms andoptionally substituted by groups R_(3a) or phenyl optionally substitutedby R_(3a);

[0038] each R_(3a) independently is R_(1c), amino, halo, cyano, nitro,thiol, alkylthio, alkylsulphonyl, alkylsulphenyl, triazolyl, imidazolyl,tetrazolyl, hydrazido, alkyl imidazolyl, thiazolyl, alkyl thiazolyl,alkyl oxazolyl, oxazolyl, alkylsulphonamido, alkylaminosulphonyl,aminosulphonyl, haloalkoxy and haloalkyl;

[0039] R_(1b), R_(1c) and R_(1j) are as defined for R_(1a);

[0040] and -L-Lp(D)_(n) is of the formula:

[0041] in which R_(r) is —(CH₂)_(c)—R_(c), —CHR_(e)R_(f),—CH₂—CHR_(e)R_(f), or R_(g) in which c is 1 or 2; R_(c) is pyridyl orphenyl (which phenyl may bear a fluoro, chloro, methyl, CONH₂, SO₂NH₂,methylaminosulphonyl, dimethylaminosulphonyl, methoxy or methylsulfonylsubstituent); each of R_(e) and R_(f) independently is hydrogen orC₁₋₃alkyl; or CHR_(e)R_(f) is cyclopentyl (which may bear a methyl,ethyl or hydroxymethyl substituent at the 3- or 4-position), cyclohexyl(which may bear a methyl, ethyl or hydroxymethyl substituent at the 3-or 4-position), tetrahydropyran-4-yl, tetrahydrothiopyran-4-yl,pyrrolidin-3-yl (which may bear a 1-methyl substituent), piperidin-4-yl(which may bear a 1-methyl substituent), or indan-2-yl; and R_(g) is2-methylsulphonylphenyl which may bear a 4-fluoro substituent or R_(g)is λ⁶-1,1-dioxobenzo[b]thiophen-7-yl;

[0042] or a physiologically-tolerable salt thereof; provided that Lp(D)nis not of the formula (K):

[0043] wherein X₂ is fluoro or hydrogen.

[0044] In the compounds of the invention, where the alpha atom is carbonit preferably has the conformation that would result from constructionfrom a D-α-aminoacid NH₂—CR_(1b)(Cy)-COOH where the NH₂ represents partof X—X. Likewise the fourth substituent R_(1b) at an alpha carbon ispreferably a methyl or hydroxymethyl group or hydrogen. It will beappreciated that the compounds of formula (I) may exist in racemic orchiral form, and that the preferred D-isomer may be administered in aracemic mixture with the L-isomer, or alone.

[0045] In the compounds of the invention, unless otherwise indicated,aryl groups preferably contain 5 to 10 ring atoms optionally including1, 2 or 3 heteroatoms selected from O, N and S; alkyl, alkenyl oralkynyl groups or alkylene moieties preferably contain up to 6 carbons,e.g. C₁₋₆ or C₁₋₃; cyclic groups preferably have ring sizes of 3 to 8atoms; and fused multicyclic groups preferably contain 8 to 16 ringatoms.

[0046] Examples of particular values for R_(1a) are: hydrogen, methyl orethyl. R_(1a) is preferably a hydrogen atom.

[0047] The linker group (X—X) from the R₂ group to the alpha atom ispreferably selected from —CH═CH—, —CONH—, —CONR_(1a)—, —NH—CO—,—NH—CH₂—, —CH₂—NH—, —CH₂O—, —OCH₂—, —COO—, —OC═O— and —CH₂CH₂—.Preferably, the X moiety nearest to the alpha atom is an NH or O atom,most preferably a NH group. The X moiety alpha to the aromatic ring ispreferably a carbon based group such as CH₂ or CO, preferably CO. Thus aparticularly preferred linker X—X is —CONH—. In an alternativeembodiment the linker is a —OCH₂— group.

[0048] Examples of particular values for R_(1b) are: hydrogen,(1-4C)alkyl, such as methyl or hydroxy(1-4C)alkyl, such ashydroxymethyl. R_(1b) is preferably a hydrogen atom.

[0049] The alpha atom (Y) is preferably a CH or C(CH₃) group. Especiallythe alpha atom (Y) is CH.

[0050] Examples of particular values for —CHR_(e)R_(f) in a—CHR_(e)R_(f), —CH₂—CHR_(e)R_(f) or —CH₂—CH₂—CHR_(e)R_(f) group are2-propyl, 3-pentyl, cyclopentyl, cyclohexyl, 4-methylcyclohexyl,tetrahydrothio-pyran-4-yl, pyrrolidin-3-yl, 1-methylpyrrolidin-3-yl,1-(2-propyl)pyrrolidin-3-yl, piperidin-4-yl, 1-methylpiperidin-4-yl,1-(2-propyl)piperidin-4-yl and indan-2-yl.

[0051] When Rr is of the formula —CHR_(e)R_(f) a preferred value for Rris 1-methylpiperidin-4-yl.

[0052] Preferably R_(r) is of the formula —(CH₂)_(c)—R_(c).

[0053] Preferably c is 2.

[0054] Preferably -L-Lp(D)_(n) is of the formula:

[0055] in which R_(r) is —(CH₂)_(c)—R_(c); in which c is 2; R_(c) isthienyl, thiazolyl (which may bear an amino substituent), isothiazolyl,oxazolyl, isoxazolyl, pyrazolyl, imidazolyl, pyridyl (which may bear anamino, methoxycarbonyl, carboxy, fluoro, cyano, methyl, methylsulphonyl,aminosulphonyl, methylaminosulphonyl or dimethylaminosulphonyl ortrifluoromethyl substituent), pyrimidinyl, pyridazinyl, pyrazinyl orphenyl (which phenyl may bear a fluoro, chloro, cyano, methyl, amino,methylsulphonyl, aminosulphonyl, methylaminosulphonyl,dimethylaminosulphonyl, methylamino, dimethylamino, carboxy,methoxycarbonyl or methoxy substituent).

[0056] Preferably, Rc is thiazolyl, (which may bear an aminosubstituent), pyrazolyl, imidazolyl, pyridyl (which may bear amethylsulphonyl, aminosulphonyl, methylaminosulphonyl,dimethylaminosulphonyl, fluoro, cyano, methyl or trifluoromethylsubstituent), pyrimidinyl, pyridazinyl, pyrazinyl or phenyl (whichphenyl may bear a fluoro, chloro, cyano, methyl, amino, methylamino,dimethylamino, carboxy, methoxycarbonyl, methylsulphonyl,aminosulphonyl, methylaminosulphonyl, dimethylaminosulphonyl or methoxysubstituent).

[0057] More preferably, Rc is thiazolyl (which may bear an aminosubstituent), pyrazolyl, imidazolyl, pyridyl (which may bear a fluoro,cyano, methyl or trifluoromethyl substituent), pyridazinyl or pyrazinyl.

[0058] Yet more preferably Rc is thiazol-2-yl, 2-aminothiazol-4-yl,pyrazol-1-yl, pyrazol-4-yl, pyridazin-3-yl, imidazol-1-yl,imidazol-4-yl, pyrazin-2-yl, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl,3-fluoropyrid-4-yl, 2-cyanopyrid-4-yl, 2-methylpyrid-4-yl or2-trifluoromethylpyrid-6-yl.

[0059] Yet more preferably, Rc is pyrazolyl, imidazolyl, pyridyl,pyridazinyl or pyrazinyl.

[0060] Preferably R_(c) is pyrid-2-yl, pyrid-3-yl or pyrid-4-yl.

[0061] Most preferably, L is CO and the lipophilic group -Lp(D)n isselected from the formulae:

[0062] wherein;

[0063] m represents 0 or 1;

[0064] X⁰ represents CH or N; and

[0065] R₃ is as defined for R3a.

[0066] Preferably m is 1.

[0067] Examples of particular values for R₃ are:—

[0068] hydrogen;

[0069] hydroxyl;

[0070] for alkoxy: methoxy or ethoxy;

[0071] for alkyl optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl: (1-6C)alkyl, such as methyl, ethyl, propyl,2-propyl, butyl, 2-butyl, t-butyl, pentyl, 2-pentyl or 3-pentyl,(1-6C)alkylamino(1-6C)alkyl, such as isopropylaminomethyl,dimethylamino-methyl, diethylaminomethyl or dimethylaminoethyl, or(1-6C)alkanoyl, such as acetyl; for hydroxyalkyl optionally substitutedby hydroxy, alkylamino, alkoxy, oxo, aryl or cycloalkyl:(1-6C)hydroxyalkyl, such as hydroxymethyl or hydroxyethyl, carboxy orcarboxy(1-5C)alkyl;

[0072] for alkoxyalkyl: methoxymethyl;

[0073] for alkoxycarbonyl: methoxycarbonyl or ethoxycarbonyl:

[0074] for alkylaminocarbonyl: methylaminocarbonyl ordimethylaminocarbonyl;

[0075] for aminoalkyl optionally substituted by hydroxy, alkylamino,alkoxy, oxo, aryl or cycloalkyl: aminomethyl, aminocarbonyl oraminocarbonyl(1-5C)alkyl;

[0076] for alkylamino optionally substituted by hydroxy, alkylamino,alkoxy, oxo, aryl or cycloalkyl: methylamino, dimethylamino, ethylamino,formylamino or acetylamino;

[0077] amino;

[0078] for halo: fluoro or chloro;

[0079] cyano;

[0080] nitro;

[0081] thiol;

[0082] for alkylthio: methylthio;

[0083] for alkylsulphonyl: methylsulphonyl, ethylsulphonyl orisopropylsulphonyl;

[0084] for alkylsulphenyl: methylsulphenyl (CH₃SO);

[0085] for triazolyl: 1,2,4-triazol-2-yl, 1,2,4-triazol-4-yl or1,2,3-triazol-4-yl;

[0086] for imidazolyl: 1,3-imidazol-1-yl or 1,3-imidazol-4-yl;

[0087] for tetrazolyl: tetrazol-1-yl or tetrazol-5-yl;

[0088] for alkylsulphonamido: methylsulphonamido, ethylsulphonamido orpropylsulphonamido;

[0089] for alkylaminosulphonyl: methylaminosulphonyl,ethylaminosulphonyl or propylaminosulphonyl; aminosulphonyl;

[0090] for haloalkoxy: trifluoromethoxy; and

[0091] for haloalkyl: trifluoromethyl or trichloromethyl.

[0092] When R₃ is present as a substituent on an aromatic ring, it ispreferably selected from hydrogen, alkylsulphonyl, aminosulphonyl,alkylaminosulphonyl, alkylaminocarbonyl, amino, amido, alkoxycarbonyl,acetylamino, chloro, fluoro, cyano, methoxy, ethoxy, nitro, hydroxy,alkylsulphonylamino, triazolyl and tetrazolyl.

[0093] When R₃ is present as a substituent on a saturated ring, it ispreferably selected from hydrogen, hydroxy, amino, (1-3C)alkoxy,(1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy, methoxycarbonyl andethoxycarbonyl.

[0094] For example specific Lp(D)n groups include

[0095] wherein R_(i) is hydrogen or (1-6C)alkyl.

[0096] Preferably Ri is hydrogen, methyl or ethyl.

[0097] More preferably Ri is hydrogen or methyl.

[0098] The cyclic group (Cy) attached to the alpha carbon is preferablyan optionally R_(3a) substituted: phenyl, pyridyl, thienyl, thiazolyl,naphthyl, piperidinyl, furanyl, pyrrolyl, isoxazolyl, isothiazolyl,pyrazolyl, oxazolyl, imidazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,pyrimidinyl, pyridazinyl, quinolyl, isoquinolyl, benzofuryl,benzothienyl or cycloalkyl group, or a phenyl group substituted byR_(3i)X_(i) in which X_(i) is a bond, O, NH or CH₂ and R_(3i) is phenyl,pyridyl or pyrimidyl group optionally substituted by R_(3a).

[0099] The cyclic group (Cy) attached to the alpha carbon is morepreferably an optionally R_(3a) substituted phenyl, pyridyl (such aspyrid-2-yl, pyrid-3-yl or pyrid-4-yl), thienyl (such as thien-2-yl orthien-3-yl), thiazolyl (such as thiazol-2-yl, thiazol-4-yl orthiazol-5-yl), naphthyl (such as naphth-1-yl), piperidinyl (such aspiperidin-4-yl) or cycloalkyl, such as a cyclohexyl group.

[0100] Examples of particular values for R_(3a) are:—

[0101] hydrogen;

[0102] hydroxyl;

[0103] for alkoxy optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl: alkoxy, such as methoxy or ethoxy, oraralkyloxy, such as benzyloxy;

[0104] for alkyl optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl: alkyl, such as methyl or ethyl, oralkylaminoalkyl, such as methylaminomethyl or dimethylaminomethyl;

[0105] for hydroxyalkyl optionally substituted by hydroxy, alkylamino,alkoxy, oxo, aryl or cycloalkyl: hydroxymethyl or carboxy;

[0106] for alkoxyalkyl: methoxymethyl;

[0107] for alkoxycarbonyl: methoxycarbonyl or ethoxycarbonyl;

[0108] for alkylaminocarbonyl: methylaminocarbonyl ordimethylaminocarbonyl;

[0109] for aminoalkyl optionally substituted by hydroxy, alkylamino,alkoxy, oxo, aryl or cycloalkyl: aminomethyl, CONH₂ or CH₂CONH₂;

[0110] for alkylamino optionally substituted by hydroxy, alkylamino,alkoxy, oxo, aryl or cycloalkyl: (1-6C)alkanoylamino, such asacetylamino;

[0111] for alkoxycarbonylamino: methoxycarbonylaminno,ethoxycarbonylamino or t-butoxycarbonylamino;

[0112] amino;

[0113] for halo: fluoro or chloro;

[0114] cyano;

[0115] nitro;

[0116] thiol;

[0117] for alkylthio: methylthio;

[0118] for alkylsulphonyl: methylsulphonyl or ethylsulphonyl;

[0119] for alkylsulphenyl: methylsulphenyl;

[0120] for alkylsulphonamido: methylsulphonylamido orethylsulphonylamido;

[0121] for alkylaminosulphonyl: methylaminosulphonyl orethylaminosulphonyl;

[0122] aminosulphonyl;

[0123] for haloalkoxy: trifluoromethoxy;

[0124] for haloalkyl: trifluoromethyl;

[0125] for a group of the formula —C(X³)N(R¹¹)R¹² (wherein X³ is O or Sand R¹¹ and R¹² are independently selected from hydrogen, methyl, ethyl,or together with the nitrogen atom to which they are attached form apyrrolidin-1-yl, piperidin-1-yl or morpholino group: —CONH₂, —CONHMe,—CON(Me)₂, —C(S)NH₂, —C(S)NHMe, —C(S)N(Me)₂,pyrrolidin-1-ylcarbonylpiperidin-1-ylcarbonyl or morpholinocarbonyl; and

[0126] —OCH₂O— which is bonded to two adjacent ring atoms in Cy.

[0127] In another aspect R_(3a) is selected from hydrogen, hydroxyl,alkoxy, alkyl (optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl), hydroxyalkyl (optionally substituted byhydroxy, alkylamino, alkoxy, oxo, aryl or cycloalkyl), alkoxyalkyl,alkoxycarbonyl, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino(optionally substituted by hydroxy, alkylamino, alkoxy, oxo, aryl orcycloalkyl), aminoalkyl (substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl), amino, halo, cyano, nitro, thiol, alkylthio,alkylsulphonyl, alkylsulphenyl, alkylsulphonamido, alkylaminosulphonyl,aminosulphonyl, haloalkoxy and haloalkyl.

[0128] Preferably X³ is O.

[0129] Examples of more specific values for R_(3a) include hydrogen,hydroxyl, methoxy, ethoxy, methyl, ethyl, hydroxymethyl, carboxy,methoxymethyl, methoxycarbonyl, ethoxycarbonyl, methylaminocarbonyl,dimethylamino-carbonyl, aminomethyl, CONH₂, CH₂CONH₂, acetylamino,methoxycarbonylamino, ethoxycarbonylamino, t-butoxycarbonylamino, amino,fluoro, chloro, bromo, cyano, nitro, thiol, methylthio, methylsulphonyl,ethylsulphonyl, methylsulphenyl, methylsulphonylamido,ethylsulphonylamido, methylaminosulphonyl, ethylaminosulphonyl,aminosulphonyl, trifluoromethoxy, trifluoromethyl, bromo, —OCH₂O— (whichis bonded to two adjacent ring atoms in Cy) and —C(X³)N(R¹¹)R¹² (whereinX³ is O or S and R¹¹ and R¹² are independently selected from hydrogen,methyl or ethyl or together with the nitrogen atom to which they areattached form a pyrrolidin-1-yl, piperidin-1-yl or morpholino group).

[0130] More examples of specific values for R_(3a) include hydrogen,hydroxyl, methoxy, ethoxy, methyl, ethyl, hydroxymethyl, carboxy,methoxymethyl, methoxycarbonyl, ethoxycarbonyl, methylaminocarbonyl,dimethylamino-carbonyl, aminomethyl, CONH₂, CH₂CONH₂, acetylamino,methoxycarbonylamino, ethoxycarbonylamino, t-butoxycarbonylamino, amino,fluoro, chloro, cyano, nitro, thiol, methylthio, methylsulphonyl,ethylsulphonyl, methylsulphenyl, methylsulphonylamido,ethylsulphonylamido, methylaminosulphonyl, ethylaminosulphonyl,aminosulphonyl, trifluoromethoxy and trifluoromethyl.

[0131] Preferably R_(3a) is hydrogen, hydroxyl, methoxy, methyl, amino,fluoro, chloro, ethylsulphonylamino, amido or methylaminocarbonyl.

[0132] Preferably Cy is selected from:

[0133] wherein:

[0134] X′ is selected from O, S and NMe;

[0135] X″ is selected from O and S;

[0136] X′″ is selected from O, S, NH and NMe;

[0137] Y′ is selected from hydrogen, amino and methyl;

[0138] R_(o) is selected from hydrogen, methyl, fluoro, chloro,trifluoromethyl, methoxy, methylthio, methylsulphinyl andmethylsulphonyl;

[0139] R_(m) is selected from hydrogen, methyl, fluoro, chloro,trifluoromethyl, methoxy, methylthio, methylsulphinyl, methylsuphonyl,carboxy, methoxycarbonyl and a group of the formula —C(X³)N(R¹¹)R¹²(wherein X³ is O or S and R¹¹ and R¹² are independently selected fromhydrogen, methyl or ethyl or

[0140] together with the nitrogen atom to which they are attached form apyrrolidin-1-yl, piperidin-1-yl or morpholino group);

[0141] R_(p) is selected from hydrogen and fluoro; or

[0142] R_(o) and R_(m) or R_(m) and R_(p) form an —OCH₂O— group; or

[0143] R_(o) and R_(m) together with the ring to which they are attachedform a 5 or 6 membered aryl or heteroaryl ring (wherein the heteroaryring contains 1 or 2 heteroatoms selected from nitrogen, oxygen andsufur);

[0144] one of R_(o1) and R_(o2) is hydrogen and the other is R_(o).

[0145] More preferably Cy is selected from phenyl (optionallysubstituted by methyl, ethyl, prop-2-yl, phenoxy, hydroxy, ethoxy,benzyloxy, prop-2-yloxy, nitro, amino, acetylamino, methylsulfonylamino,dimethylamino, chloro, methoxy, trifluoromethyl, methylthio,methylsulfonyl, tert-butylthio, tert-butylsulfonyl, aminosulfonyl orcarbamoyl), pyridyl, thienyl, furanyl, imidazolyl, thiazolyl (optionallysubstituted by amino), napththyl, isoquinolinyl and quinolinyl.

[0146] Yet more preferably, Cy is selected from phenyl, 2-chlorophenyl,2-methoxyphenyl, 4-carbamoylphenyl, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl,thien-2-yl, thien-3-yl, furan-2-yl, furan-3-yl, imidazol-2-yl,thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, naphthyl, isoquinolin-5-yl,isoquinolin-8-yl, quinolin-4-yl, quinolin-5-yl, and quinolin-8-yl.

[0147] Other examples of values for Cy are 4-aminophenyl,4-N-methylamidophenyl, 4-(N,N-dimethyl)amidophenyl, 2-methylphenyl,2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 4-hydroxyphenyl,4-methoxyphenyl, 4-carboxyphenyl, 3-ethylsulphonylaminophenyl,2-methylthiazol-4-yl, 1-ethylpiperidin-4-yl, cyclopentyl, cyclohexyl,naphth-1-yl, 2-aminothiazol-4-yl, 2-trifluoromethylphenyl,3-methylthiophenyl, 2-methylsulphonylphenyl, 3-bromophenyl,3-cyanophenyl and benzo[b]thiophen-3-yl.

[0148] Yet more preferably Cy is selected from phenyl, 2-chlorophenyl,2-methoxyphenyl, 4-carbamoylphenyl, pyrid-2-yl, pyrid-4-yl, thien-2-yl,thien-3-yl, furan-2-yl, furan-3-yl, imidazol-2-yl, thiazol-2-yl,thiazol-4-yl, thiazol-5-yl and quinolin-4-yl.

[0149] Most preferably, Cy is selected from phenyl, 2-methoxyphenyl,4-carbamoylphenyl and pyrid-2-yl.

[0150] Most preferably Cy is phenyl.

[0151] Examples of particular values for R_(1c) are:

[0152] hydrogen;

[0153] hydroxyl;

[0154] for alkoxy: methoxy or ethoxy;

[0155] for alkyl optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl: alkyl, such as methyl or ethyl, oralkylaminoalkyl, such as methylaminomethyl or dimethylaminomethyl;

[0156] for hydroxyalkyl: hydroxymethyl; for alkoxyalkyl: methoxymethyl;

[0157] for alkoxycarbonyl: methoxycabonyl or ethoxycarbonyl;

[0158] for alkylaminocarbonyl: methylaminocarbonyl ordimethylaminocarbonyl;

[0159] for alkoxycarbonylamino: methoxycarbonylamino,ethoxycarbonylamino or t-butoxycarbonylamino;

[0160] for alkylamino optionally substituted by hydroxy, alkylamino,alkoxy, oxo, aryl or cycloalkyl: (1-6C)alkanoylamino, such asacetylamino; and

[0161] for aminoalkyl substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl: aminomethyl, CONH₂ or CH₂CONH₂.

[0162] Referring to R², examples of a 5 or 6 membered aromatic carbonring optionally interrupted by a nitrogen, oxygen or sulphur ring atomin R² are phenyl; pyrrolyl, such as 2-pyrrolyl; pyridyl, such as3-pyridyl; pyrazinyl, such as 2-pyrazinyl; furyl, such as 2-furyl; andthienyl, such as 2-thienyl or 3-thienyl. Preferably the ring isinterrupted (i.e. a carbon atom is replaced) by at most one heteroatom.In another aspect the ring is phenyl, 2-thienyl or 2-pyrrolyl. In yetanother aspect, the ring is phenyl.

[0163] When the ring is phenyl, the group R₂ may be a group of formula

[0164] in which R₅ is amino, hydroxy or hydrogen, and R₆ and R₇ whichmay be the same or different represent halo, nitro, thiol, cyano,haloalkyl, haloalkoxy, amido, hydrazido, amino, alkylthio, alkenyl,alkynyl or R₁ or taken together form a 5 or 6 membered fused carbocyclicring or 5 membered heterocyclic ring, which may itself be substituted byR_(1j), amino, halo, cyano, nitro, thiol, alkylthio, haloalkyl,haloalkoxy.

[0165] When the substituents at the 3 and 4 positions taken togetherform a fused ring which is a 5 or 6 membered carbocyclic or heterocyclicring, examples of the resultant bicyclic ring are naphthyl, such as2-naphthyl; benzimidazolyl, such as benzimidazol-5-yl orbenzimidazol-6-yl; isoquinolinyl, such as isoquinolin-7-yl; indolyl,such as indol-2-yl, indol-5-yl or indol-6-yl; indazolyl, such asindazol-5-yl; indazol-6-yl; 3,4-methylenedioxyphenyl; dihydroindolyl,such as 2,3-dihydroindol-6-yl; benzothiazolyl, such as benzothiazol-2-ylor benzothiazol-6-yl; benzo[b]thiophenyl, such as benzo[b]thiophen-2-yl;benzofuryl, such as benzofur-2-yl; imidazo[1,2-a]pyrimidinyl, such asimidazo[1,2-a]pyrimidin-2-yl; tetrahydroimidazo[1,2-a]pyrimidinyl, suchas tetrahydroimidazo[1,2-a]pyrimidin-2-yl; and benzisoxazolyl, such asbenzisoxazol-5-yl.

[0166] Preferably, R₂ is phenyl, thien-2-yl, naphthyl, indol-2-yl,indol-6-yl, benzo[b]furan-5-yl, benzo[b]thiophen-2-yl orbenzimidazol-2-yl (each of which is optionally substituted ashereinabove defined).

[0167] It is preferred that at least one of R₆ and R₇ be other thanhydrogen and that R₆, if present, is preferably a substituent containingone or more polar hydrogens such as hydroxy, amino, alkylamino,alkylaminoalkyl, aminocarbonyl, alkylaminocarbonyl, hydrazo andalkylhydrazo; alternatively R₆ and R₇ are joined together in theformation of a naphthyl or indolyl or azaindolyl or diazaindolyl group.

[0168] It is especially preferred that R₆ be amino and R₇ be chloro,bromo, methyl, methoxy or vinyl; or that R₆ and R₇ taken together forman indolyl ring with the NH at the 6-position or taken together form anaphthyl ring.

[0169] In another aspect R₂ represents:

[0170] (i) phenyl optionally being substituted in the 3 and/or 4position by halo, nitro, thiol, haloalkoxy, hydrazido, alkylhydrazido,amino, cyano, haloalkyl, alkylthio, alkenyl, alkynyl, acylamino, tri ordifluoromethoxy, carboxy, acyloxy, MeSO₂— or R₁, and optionallysubstituted at the 6 position by amino, hydroxy, halo, alkyl, carboxy,alkoxycarbonyl, cyano, amido, aminoalkyl, alkoxy or alkylthio;

[0171] (ii) naphth-2-yl optionally substituted at the 6 or 7 position byhalo, haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R_(1j) and optionally substituted at the 3 positionby amino, hydroxy, halo, alkyl, carboxy, cyano, amido, aminoalkyl,alkoxy or alkylthio;

[0172] (iii) isoquinolin-7-yl, indol-5-yl, indol-6-yl, indazol-5-yl,indazol-6-yl, benzothiazol-6-yl or benzisoxazol-5-yl optionallysubstituted at the 3 position by halo, haloalkoxy, haloalkyl, cyano,nitro, amino, hydrazido, alkylthio, alkenyl, alkynyl or R_(1j);

[0173] (iv) benzimidazol-5-yl or benzothiazol-6-yl optionallysubstituted at the 2 position by amino;

[0174] (v) thien-2-yl or thien-3-yl optionally substituted at the 4 or 5position by halo, haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido,alkylthio, alkenyl, alkynyl or R₁;

[0175] (vi) 3,4-methylenedioxyphenyl, 2,3-dihydroindol-6-yl,3,3-dichloro-2-oxo-indol-6-yl or 1-methyl-3-aminoindazol-5-yl;

[0176] (vii) benzothiazol-2-yl, imidazo[1,2-a]pyrimidin-2-yl ortetrahydroimidazo[1,2-a]pyrimidin-2-yl;

[0177] (viii) pyrazol-2-yl optionally substituted at the 5 position byhalo, haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R₁;

[0178] (ix) pyrid-2-yl optionally substituted at the 5 position by halo,haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R₁;

[0179] (x) pyrid-3-yl optionally substituted at the 6 position by halo,haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R₁;

[0180] (xi) benzofur-2-yl optionally substituted at the 3 position byamino, hydroxy, halo, alkyl, carboxy, cyano, amido, aminoalkyl, alkoxyor alkylthio and at the 5 or 6 position by halo, haloalkoxy, haloalkyl,cyano, nitro, amino, hydrazido, alkylthio, alkenyl, alkynyl or R_(1j);

[0181] (xii) indol-2-yl optionally substituted on the indole nitrogenatom by alkyl and optionally substituted at the 5 or 6 position by halo,haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R_(1j);

[0182] (xiii) indol-6-yl substituted at the 5 position by amino,hydroxy, halo (such as fluoro or chloro), alkyl, carboxy,alkoxycarbonyl, cyano, amido, aminoalkyl, alkoxy or alkylthio andoptionally substituted at the 3 position by halo (such as chloro),haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R_(1j); or

[0183] (xiv) benzo[b]thiophen-2-yl optionally substituted at the 3position by amino, hydroxy, halo, alkyl, carboxy, cyano, amido,aminoalkyl, alkoxy or alkylthio and at the 5 or 6 position by halo,haloalkoxy, haloalkyl, cyano, nitro, amino, hydrazido, alkylthio,alkenyl, alkynyl or R_(1j).

[0184] Examples of particular values for substituents that may bepresent on R₂ are:

[0185] for halo: fluoro, chloro, bromo or iodo;

[0186] nitro;

[0187] thiol;

[0188] for haloalkoxy: difluoromethoxy or trifluoromethoxy;

[0189] hydrazido;

[0190] for alkylhydrazido: methylhydrazido;

[0191] amino;

[0192] cyano;

[0193] for haloalkyl: trifluoromethyl;

[0194] for alkylthio: methylthio;

[0195] for alkenyl: vinyl;

[0196] for alkynyl: ethynyl;

[0197] for acylamino: acetylamino;

[0198] carboxy;

[0199] for acyloxy: acetoxy;

[0200] hydroxy;

[0201] for alkyl: methyl or ethyl;

[0202] amido (CONH₂);

[0203] for aminoalkyl: aminomethyl; and

[0204] for alkoxy: methoxy or ethoxy.

[0205] Preferably R₂ is optionally substituted by 1 or 2 substituentsselected from fluoro, chloro, amino, methyl, ethyl and methoxy.

[0206] Examples of particular values for R₁ are:

[0207] hydrogen;

[0208] hydroxy;

[0209] for alkoxy: methoxy or ethoxy;

[0210] for alkyl optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl: alkyl, such as methyl or ethyl,alkylaminoalkyl, such as dimethylaminomethyl, or alkanoyl, such asacetyl;

[0211] for hydroxyalkyl: hydroxymethyl;

[0212] for alkoxyalkyl: methoxymethyl;

[0213] for alkoxycarbonyl: methoxycarbonyl;

[0214] for alkylaminocarbonyl: methylaminocarbonyl;

[0215] for alkylamino: methylamino, ethylamino or dimethylamino;

[0216] for hydroxyalkyl substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl: carboxyl or carboxymethyl; and

[0217] for aminoalkyl substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl: amido (CONH₂) or amidomethyl.

[0218] Examples of particular values for R_(1j) are:

[0219] hydrogen;

[0220] hydroxy;

[0221] for alkoxy: methoxy or ethoxy;

[0222] for alkyl optionally substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl: alkyl, such as methyl or ethyl, or alkanoyl,such as acetyl;

[0223] for hydroxyalkyl: hydroxymethyl;

[0224] for alkoxyalkyl: methoxymethyl;

[0225] for alkoxycarbonyl: methoxycarbonyl;

[0226] for alkylamino: methylamino, ethylamino or dimethylamino;

[0227] for hydroxyalkyl substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl: carboxyl or carboxymethyl; and

[0228] for aminoalkyl substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl: amido (CONH₂) or amidomethyl.

[0229] In yet another aspect R₂ represents:

[0230] (i) phenyl optionally being substituted in the 3 and/or 4position by fluoro, chloro, bromo, iodo, nitro, difluoromethoxy,trifluoromethoxy, amino, cyano, trifluoromethyl, methylthio, vinyl,carboxy, acetoxy, MeSO₂—, hydroxy, methoxy, ethoxy, methyl,methoxycarbonyl, methylamino, ethylamino or amido, and optionallysubstituted at the 6 position by amino, hydroxy, fluoro,methoxycarbonyl, cyano or aminomethyl (preferably phenyl substituted inthe 4 position by chloro, amino, vinyl, methylamino, methyl or methoxy,optionally at the 3 position with amino or hydroxy, and optionally atthe 6 position with amino or hydroxy);

[0231] (ii) naphth-2-yl optionally substituted at the 6, position byhydroxy and optionally substituted at the 3 position by amino orhydroxy;

[0232] (iii) isoquinolin-7-yl, indol-5-yl, indol-6-yl, indazol-5-yl,indazol-6-yl, benzothiazol-6-yl or benzisoxazol-5-yl optionallysubstituted at the 3 position by chloro, bromo, amino, methyl or methoxy(preferably indol-6-yl optionally substituted at the 3 position bychloro, bromo, methyl or methoxy);

[0233] (iv) benzimidazol-5-yl or benzothiazol-6-yl optionallysubstituted at the 2 position by amino;

[0234] (v) thien-2-yl or thien-3-yl optionally substituted at the 4 or 5position by methylthio, methyl or acetyl;

[0235] (vi) 3,4-methylenedioxyphenyl, 2,3-dihydroindol-6-yl,3,3-dichloro-2-oxo-indol-6-yl or 1-methyl-3-aminoindazol-5-yl;

[0236] (vii) benzothiazol-2-yl, imidazo[1,2-a]pyrimidin-2-yl ortetrahydroimidazo[1,2-a]pyrimidin-2-yl;

[0237] (viii) pyrazol-2-yl substituted at the 5 position by methyl;

[0238] (ix) pyrid-2-yl optionally substituted at the 6 position bychloro;

[0239] (x) pyrid-3-yl optionally substituted at the 4 position bychloro;

[0240] (xi) benzofur-2-yl optionally substituted at the 3 position bychloro, methyl or methoxy, at the 5 or 6 position by methyl and at the 6position by methoxy;

[0241] (xii) indol-2-yl optionally substituted on the indole nitrogenatom by methyl and optionally substituted at the 5 or 6 position byfluoro, chloro, bromo, methyl or methoxy;

[0242] (xiii) indol-6-yl substituted at the 5 position by chloro, fluoroor hydroxy and optionally substituted at the 3 position by chloro ormethyl; or

[0243] (xiv) benzo[b]thiophen-2-yl optionally substituted at the 3position by fluoro, chloro or methyl, and optionally substituted at the5 or 6 position by fluoro, chloro, methyl, hydroxy, or methoxy.

[0244] Particular values for R₂ are:

[0245] (i) phenyl, 2-aminophenyl, 3-aminophenyl, 2-amino-3-fluorophenyl,2-amino-4-fluorophenyl, 2-amino-4-chlorophenyl, 2-amino-3-bromophenyl,2-amino-3-nitrophenyl, 2-amino-4-nitrophenyl,3,4-dimethoxy-5-aminophenyl, 2-amino-4-methylphenyl,2-amino-3-methylphenyl, 2-amino-3-methoxyphenyl, 3,4-diaminophenyl,3,5-diaminophenyl, 3-amino-4-fluorophenyl, 3-amino-4-chlorophenyl,3-amino-4-bromophenyl, 3-amino-4-hydroxyphenyl,3-amino-4-carboxymethylphenyl, 3-amino-4-methylphenyl,3-amino-4-methoxyphenyl, 2-fluorophenyl, 4-fluoro-3-cyanophenyl,3-chlorophenyl, 3-chloro-4-hydroxphenyl, 3-chloro-5-hydroxyphenyl,4-chlorophenyl, 4-chloro-2-hydroxyphenyl, 4-chloro-3-hydroxyphenyl,4-chloro-3-methylphenyl, 4-chloro-3-methoxyphenyl, 4-bromophenyl,4-bromo-3-methylphenyl, 4-iodophenyl, 2-cyanophenyl, 3-cyanophenyl,4-cyanophenyl, 3-cyano-5-aminophenyl, 2-hydroxphenyl,2-hydroxy-4-methoxyphenyl, 3-hydroxphenyl, 3-hydroxy-4-methylphenyl,2,4-dihydroxyphenyl, 3,4-dihydroxyphenyl, 3-hydroxy-4-methoxyphenyl,4-difluoromethoxyphenyl, 4-trifluoromethoxphenyl,4-trifluoromethylphenyl, 4-methylthiophenyl, 4-ethoxycarbonylphenyl,4-acetoxyphenyl, 4-ethanesulfonylphenyl, 3-methylphenyl,3-methyl-5-aminophenyl, 4-methylphenyl, 4-vinylphenyl, 4-methoxyphenyl,4-ethoxyphenyl, 4-methoxy-3-chlorophenyl, 4-methoxy-3-methylphenyl,3-methylaminophenyl, 4-methylaminophenyl, 4-ethylaminophenyl or2-aminomethylphenyl;

[0246] (ii) naphth-2-yl, 3-aminonaphth-2-yl, 3-hydroxynaphth-2-yl or6-hydroxynaphth-2-yl;

[0247] (iii) isoquinolin-7-yl, indol-5-yl, indol-6-yl,3-chloroindol-6-yl, 3-bromoindol-6-yl, 3-methylindol-6-yl,3-methoxyindol-6-yl, indazol-5-yl, 3-aminoindazol-5-yl, indazol-6-yl,benzothiazol-6-yl, 3-aminobenzisoxazol-5-yl;

[0248] (iv) benzimidazol-5-yl, 2-aminobenzimidazol-5-yl, orbenzothiazol-6-yl;

[0249] (v) thien-2-yl, 5-methylthien-2-yl, 5-methylthio-thien-2-yl,5-acetylthien-2-yl or thien-3-yl;

[0250] (vi) 3,4-methylenedioxyphenyl, 2,3-dihydroindol-6-yl,3,3-dichloro-2-oxo-indol-6-yl or 1-methyl-3-aminoindazol-5-yl;

[0251] (vii) benzothiazol-2-yl, imidazo[1,2-a]pyrimidin-2-yl ortetrahydroimidazo[1,2-a]pyrimidin-2-yl;

[0252] (viii) 5-methylpyrazol-2-yl;

[0253] (ix) 5-chloropyrid-2-yl;

[0254] (x) pyrid-3-yl, 6-chloropyrid-3-yl;

[0255] (xi) benzofur-2-yl, 5-chlorobenzofur-2-yl, 3-methylbenzofur-2-yl,5-methylbenzofur-2-yl, 6-methoxybenzofur-2-yl;

[0256] (xii) indol-2-yl, 5-fluoroindol-2-yl, 5-chloroindol-2-yl,5-methylindol-2-yl, 5-methoxindol-2-yl, 6-methoxyindol-2-yl and1-methyl-indol-2-yl;

[0257] (xiii) 5-fluoroindol-6-yl; or

[0258] (xiv) benzo[b]thiophen-2-yl, 5-chloro-benzo[b]thiophen-2-yl or6-chlorobenzo[b]thiophen-2-yl.

[0259] Preferably, R₂ is selected from one of the formulae (A′) to (H′):

[0260] wherein X₄ is O or S, R₁₃ is selected from hydrogen, fluoro[except for (C′)], chloro or methyl and R₁₄ is selected from hydrogen,methyl, ethyl, fluoro, chloro, and methoxy and R₁₅ is selected fromhydrogen, methyl, fluoro, chloro and amino.

[0261] More preferably, R₂ is of the formula (A′) (wherein R₁₄ isselected from hydrogen, methyl, ethyl, fluoro, chloro, and methoxy andR₁₅ is selected from hydrogen, methyl, fluoro, chloro and amino) or ofthe formula (B′) (wherein R₁₃ is chloro) or of the formula (C′) (whereinR₁₃ is selected from hydrogen, methyl and chloro) or of the formula (D′)(wherein R₁₃ is selected from hydrogen, methyl, fluoro and chloro) or ofthe formula (E′) (wherein R₁₃ is hydrogen) or of the formula (G′)(wherein R₁₃ is chloro).

[0262] Yet more preferably, R₂ is 4-chlorophenyl, 4-methoxyphenyl,3-amino-4-chlorophenyl, indol-2-yl, 5-chloroindol-2-yl, indol-6-yl,3-chloroindol-6-yl or 3-methylindol-6-yl. Another R₂ group of particularinterest is 3-aminobenzisoxazol-5-yl.

[0263] Yet more preferably, R₂ is of the formula (A′) or (C′) and R₁₃,R₁₄ and R₁₅ are as defined hereinabove.

[0264] Most preferably, R₂ is of the formula (A′) and R₁₄ is methoxy andR₁₅ is hydrogen or of the formula (C′) and R₁₃ is hydrogen, methyl orchloro.

[0265] Another preferred compound of the present invention is one of theformula:

[0266] wherein Cy and R₂ are as herinabove defined.

[0267] A preferred compound of the present invention is of the formula:

[0268] wherein Cy, R₂ and R_(c) are as hereinabove defined.

[0269] Especial mention may be made of:—

[0270]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)-ethyl]piperazine;

[0271]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine;

[0272]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine;

[0273]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine;

[0274]1-(4-Methoxybenzoyl-D-(2-chlorophenyl)glycinyl)-4-(1-methylpiperidin-4-yl)piperazine;

[0275]1-(Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl)-4-(1-methylpiperidin-4-yl)piperazine;and

[0276]1-(4-Methoxybenzoyl-D-(2-trifluoromethylphenyl)glycinyl)-4-(1-methylpiperidin-4-yl)piperazine;

[0277] and physiologically-tolerable salts thereof.

[0278] Compounds in this group have been found to have good oralexposure and a desirable pharmacological/toxicological profile.

[0279] The compounds of the invention may be prepared by conventionalchemical synthetic routes or by routes as illustrated by the followingexamples.

[0280] The compounds of the formula (I) may be prepared by forming the—X—X— bond from appropriate intermediates. For example, when —X—X— is—CONH— or —CO—NR_(1a)—, by reacting a compound of the formula (10):H₂N—Y-(Cy)-L-Lp(D)_(n) with a compound of the formula R₂—COOH, underconditions known for the formation of an amide bond. The reaction isconveniently carried out in the presence of a benzotriazole-basedreagent such as 1-hydroxybenzotriazole or 1-hydroxy-7-azabenzotriazole,in an inert organic solvent such as dimethylformamide and/or methylenechloride. The reaction mixture is usually taken to 0° C. and then adehydrating agent such as dicyclohexylcarbodiimide or1-(3-dimethylaminopropyl)-3-ethylcarbodiimide added. Other suitablereagents and solvents are known in the art, for example an acid halide,such as R₂—COCl.

[0281] Compounds wherein —X—X— is —NHCO— or —NHCH₂— may be formed fromthe appropriate intermediates using reaction conditions for theformation of an amide bond as described above and if necessarysubsequent reduction of the resulting amide bond.

[0282] Compounds of the formula (I) wherein —X—X— is of the formula—CH₂NH— may be prepared by reducing the corresponding compound of theformula (I) wherein —X—X— is —CONH— or by reaction of a compound offormula (10): H₂N—Y-(Cy)-L-Lp(D)_(n) with a compound of the formulaR₂CHO and reducing the intermediate of the formula (I) wherein —X—X— is—C═N— with, for example, sodium cyanoborohydride.

[0283] When —X—X— is —CH═CH—, the compounds of the formula (I) may beprepared using the Wittig or Horner-Emmons reactions. The correspondingcompound in which —X—X— is —CH₂CH₂— can be formed by reduction of the—CH═CH— group, for example with hydrogen over a palladium-on-carboncatalyst.

[0284] An —X—X— bond of the formula —COO— or —OC(O)— may be formed byreacting the appropriate hydroxy and activated carboxylic acid (e.g.acid chloride or reactive ester) intermediates under conditions knownfor ester bond formation. Alternatively, a hydroxy and a carboxylic acidintermediate could be reacted together in the presence ofdiethylazodicarboxylate/triphenylphosphine.

[0285] An —X—X— bond of the formula —CH₂O— or —OCH₂— may be formed byreacting the appropriate hydroxy intermediate with the appropriate alkylhalide in the presence of a base. Conditions for the formation of anether bond are known in the art.

[0286] These reactions can also be used to form intermediates, whichcontain one of the above —X—X— bonds.

[0287] Compounds of the formula (I) in which R_(r) is —(CH₂)_(c)—R_(c)may also be prepared by reductive coupling a compound of the formula(11):

[0288] with a compound of formula (12)

OHC—(CH₂)_(c−1)—R_(c)

[0289] The reaction is conveniently performed in the presence of areducing agent, such as sodium cyanobrohydride. Convenient solventsinclude alcohols, such as methanol, optionally with a halogenatedhydrocarbon as solvent, such as 1,2-dichloroethane, and acetic acid. Thecoupling is conveniently effected at a temperature in the range of from0 to 100° C.

[0290] The intermediates of formula (11) are believed to be novel, andare provided as a further aspect of the invention.

[0291] The intermediates of formula (11) in which X—X is CONH may beprepared by reacting a compound of formula (13)

[0292] in which Pg¹ represents an amino protecting group, such ast-butoxycarbonyl, with a compound of formula R₂—COOH, under conditionsknown for the formation of an amide bond, for example as describedhereinabove for forming a compound of formula (I), followed bydeprotection.

[0293] The compounds of formula (13) may be prepared by reacting anappropriate N-protected glycine of formula (14)

[0294] in which Pg² represents an amino protecting group that can beselectively removed in the presence of Pg¹ (for example, when Pg¹ ist-butoxycarbonyl, Pg² may be be benzyloxycarbonyl), with a compound offormula (15)

[0295] under amide bond forming conditions, followed by selectivelyremoving the protecting group Pg².

[0296] Compounds of the formula (10) in which X is CONH may be preparedby deprotecting a compound of the formula (16):

[0297] in which Pg³ represents an amino protecting group, such ast-butoxycarbonyl.

[0298] The intermediates of formula (16) and the corresponding amineswithout Pg³ are believed to be novel, and are provided as a furtheraspect of the invention.

[0299] Compounds of formula (16) may be prepared by reacting a compoundof formula (14) with a compound of formula (17)

[0300] under amide bond forming conditions. The reaction is convenientlyperformed in the presence of diethylcyano-phosphate. Convenient solventsinclude amides, such as dimethylformamide. The temperature isconveniently in the range of from 0 to 100° C.

[0301] Compounds of formula (17) in which R_(r) is —(CH₂)_(c)—R_(c), maybe prepared by reacting a compound of formula (18)

[0302] in which Pg⁴ represents an amino protecting group, such ast-butoxycarbonyl, with a compound of formula (19)

[0303] followed by removing the protecting group, Pg⁴. The reaction isconveniently performed in the presence of an acid, such as acetic acid.Convenient solvents include alcohols, such as ethanol.

[0304] Compounds of formula (17) in which R_(r) is —(CH₂)_(c)—R_(c) mayalso be prepared by reacting a compound of formula (18) with a compoundof formula (19a)

Z-(CH₂)_(c)—R_(c)

[0305] in which Z represents a leaving atom or group, such asmethanesulfonyloxy or benzenesulfonyloxy, followed by removing theprotecting group, Pg⁴.

[0306] Compounds of formula (17) in which R_(r) is —(CH₂)_(c)—R_(c) mayalso be prepared by reducing compounds of formula (20) or formula (20A)

[0307] in which Pg⁵ and Pg⁶ each represent an amino protecting group,such as t-butoxycarbonyl, followed by removing the protecting group,Pg⁵. The reduction is conveniently performed in the presence of areducing agent, such as borane, in an ether such as tetrahydrofuran.

[0308] Compounds of formula (20) may be prepared by reacting a compoundof formula (18) with a compound of formula (21)

HOOC—(CH₂)_(c−1)—R_(c)

[0309] under amide bond forming conditions.

[0310] Alternatively, compounds of formula (20) may be prepared byreacting a compound of formula (18) with a compound of formula (21a)

HOOC—(CH)_(c−1)X′  (21a)

[0311] in which X′ is a hydrogen atom, such as bromine, followed byreaction with a compound of formula (21b)

HR_(c)

[0312] in the presence of a strong base, such as sodium hydride.

[0313] Hence the present invention also provides a process for thepreparation of a compound of formula (I) comprising:

[0314] a) when —X—X is —CONH—, reacting a compound of formula (10) witha compound of formula R₂—COOH, under amide bond-forming conditions; or

[0315] b) when R_(r) is —(CH₂)_(c)—R_(c), reacting a compound of formula(11) with a compound of (12);

[0316] wherein R₂, X, Y, Cy, c and R_(r) are as hereinabove defined andformulae (10), (11) and (12) are as hereinabove defined, followed if asalt is required, by forming a physiologically acceptable salt.

[0317] An amino acid of formula (23)

[0318] or an N-protected glycine of formula (14) may be prepared (forexample) by one or more of the following methods:

[0319] (i) from aryl or heteroaryl aldehydes via the Strecker synthesisor modifications thereof, via Bucherer-Bergs hydantoin synthesis, or viathe Ugi methodology (“Isonitrile Chemistry”, Ugi I. Ed.; Academic: NewYork, 1971;145-1999, “Multicomponent Reactions with Isocyanides”,Domling, A.; Ugi, I. Angew. Chem. Int. Ed. 2000, 39, 3168; “Amino AcidDerivatives by Multicomponent Reactions”, Dyker, G. Angew, Chem. Int.Ed. Engl. 1997, 36, 1700; and also see “A new Class of ConvertibleIsocyanides in the Ugi Four-Component Reaction”, Lindhorst, T.; Bock H.;Ugi, I. Tetrahedron, 1999, 55, 7411.) with removal and replacement ofprotecting groups;

[0320] (ii) from styrenes via Sharpless methodology (J. Am. Chem. Soc.1998,120, 1207-1217)

[0321] (iii) from aryl boronic acids via Petasis methodology(Tetrahedron, 1997, 53, 16463-16470) with removal and replacement ofprotecting groups;

[0322] (iv) from aryl and heteroaryl acetic acids—via Evan's azidation(Synthesis, 1997, 536-540) or by oximation, followed by reduction andaddition of protecting groups; or

[0323] (v) from existing aryl glycines by manipulation of functionalgroups, for example, alkylation of hydroxy groups, palladium assistedcarbonylation of triflates derived from hydroxy groups and furthermanipulation of the carboxylic esters to give carboxylic acids byhydrolysis, carboxamides by activation of the carboxylic acid andcoupling with amines, amines via Curtius reaction on the carboxylicacid;

[0324] (vi) from aliphatic, carbocylic and non-aromatic heterocyclicaldehydes and ketones using a Horner-Emmons reaction withN-benzyloxycarbonyl)-α-phosphonoglycine trimethyl ester (Synthesis,1992, 487-490); or

[0325] (vii) from oximes of formula

[0326] in which Pg is a carboxy protecting group, by reduction. (Oximesin which Cy is a heteroaryl group may be prepared from compounds offormula

[0327] Alternatively, oximes may be prepared by nitrosation of acompound of formula Cy-CH₂—COOPg, or by reaction of hydroxylamine with acompound of formula Cy-CO—COOPg).

[0328] A starting material for the preparation of a compound of formula(I), where the alpha atom is nitrogen, may be produced, for example, byreaction of a beta protected hydrazine (such protection to be chosen asto be compatible with the subsequent reagents to be employed) withphosgene, diphosgene, triphosgene or N,N′carbonyl diimidazole to give areactive compound of the type PGNHN(Cy)COCl or PGNHN(Cy)CO-imidazole(wherein PG is a protecting group).

[0329] This intermediate may be used as has been described above for thecarboxylic starting reagents where the alpha atom is carbon.

[0330] The skilled person will be aware that at certain stages in thesynthesis of a compound of formula (I) it may be necessary to protect areactive functional group in the molecule to prevent unwantedside-reactions.

[0331] The protection of amino and carboxylic acid groups is describedin McOmie, Protecting Groups in Organic Chemistry, Plenum Press, NY,1973, and Greene and Wuts, Protecting Groups in Organic Synthesis, 2nd.Ed., John Wiley & Sons, NY, 1991. Examples of carboxy protecting groupsinclude C₁-C₆ alkyl groups such as methyl, ethyl, t-butyl and t-amyl;aryl(C₁-C₄)alkyl groups such as benzyl, 4-nitrobenzyl, 4-methoxybenzyl,3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl,2,4,6-trimethylbenzyl, benzhydryl and trityl; silyl groups such astrimethylsilyl and t-butyldimethylsilyl; and allyl groups such as allyland 1-(trimethylsilylmethyl)prop-1-en-3-yl.

[0332] Examples of amine protecting groups (PG) include acyl groups,such as groups of formula RCO in which R represents C₁₋₆ alkyl, C₃₋₁₀cycloalkyl, phenyl C₁₋₆ alkyl, phenyl, C₁₋₆ alkoxy, phenyl C₁₋₆ alkoxy,or a C₃₋₁₀ cycloalkoxy, wherein a phenyl group may be optionallysubstituted, for example by one or two of halogen, C₁-C₄ alkyl and C₁-C₄alkoxy.

[0333] Preferred amino protecting groups include benzyloxycarbonyl(CBz), t-butoxycarbonyl (Boc) and benzyl.

[0334] In another aspect the invention relates to a process forpreparing a compound of formula I comprising deprotecting a compound offormula (I′):

R²′—X—X—Y(Cy′)-L-Lp(D)_(n)′  (I)′

[0335] Wherein R²′ is R² (as hereinabove defined) or protected R², Cy′is Cy (as hereinabove defined) or protected Cy and Lp(D)_(n)′ isLp(D)_(n) (as hereinabove defined) or protected Lp(D)_(n); providing atleast one protecting group is present.

[0336] If necessary physiologically tolerable salts can be formed usingmethods known in the art.

[0337] It will be understood that the compounds of formula (I) may beisolated in the form of salts or solvates (which may or may not bephysiologically tolerable), and that all such salts and solvates aretherefore included within the scope of the present invention.

[0338] All novel intermediates described herein, for example thecompounds of formula

[0339] and salts thereof, are provided as further aspects of theinvention.

[0340] The compounds of the invention may be administered by anyconvenient route, e.g. into the gastrointestinal tract (e.g. rectally ororally), the nose, lungs, musculature or vasculature or transdermally.The compounds may be administered in any convenient administrative form,e.g. tablets, powders, capsules, solutions, dispersions, suspensions,syrups, sprays, suppositories, gels, emulsions, patches etc. Suchcompositions may contain components conventional in pharmaceuticalpreparations, e.g. diluents, carriers, pH modifiers, sweeteners, bulkingagents, and further active agents. Preferably the compositions will besterile and in a solution or suspension form suitable for injection orinfusion. Such compositions form a further aspect of the invention.

[0341] The following are examples of pharmaceutical compositions ofcompounds according to the invention.

Formulation 1 Hard Gelatin Capsules are Prepared Using the FollowingIngredients

[0342] Quantity (mg/capsule) Active Ingredient 250 Starch, dried 200Magnesium stearate  10 Total 460 mg

[0343] The above ingredients are mixed and filled into hard gelatincapsules in 460 mg quantities.

Formulation 2 Tablets Each Containing 60 mg of Active Ingredient areMade as Follows

[0344] Active Ingredient   60 mg Starch   45 mg Microcrystallinecellulose   35 mg Polyvinylpyrrolidone   4 mg Sodium carboxymethylstarch  4.5 mg Magnesium stearate  0.5 mg Talc   1 mg Total  150 mg

[0345] The active ingredient, starch, and cellulose are passed through aNo. 45 mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders which are thenpassed through a No. 14 mesh U.S. sieve. The granules so produced aredried at 50° C. and passed through a No. 18 mesh U.S. sieve. The sodiumcarboxymethyl starch, magnesium stearate, and talc, previously passedthrough a No. 60 mesh U.S. sieve, are then added to the granules which,after mixing, are compressed on a tablet machine to yield tablets eachweighing 150 mg.

[0346] Viewed from this aspect the invention provides a pharmaceuticalcomposition comprising a serine protease inhibitor according to theinvention together with at least one pharmaceutically acceptable carrieror excipient. The pharmaceutical composition may also optionallycomprise at least one further antithrombotic and/or thrombolytic agent.

[0347] Viewed from a further aspect the invention provides the use of aserine protease inhibitor according to the invention for the manufactureof a medicament for use in a method of treatment of the human ornon-human animal body (e.g. a mammalian, avian or reptilian body) tocombat (i.e. treat or prevent) a condition responsive to said inhibitor.

[0348] Viewed from a further aspect the invention provides a method oftreatment of the human or non-human animal body (e.g. a mammalian, avianor reptilian body) to combat a condition responsive to a serine proteaseinhibitor (e.g. a condition such as a thrombotic disorder responsive toa factor Xa inhibitor), said method comprising administering to saidbody an effective amount of a serine protease inhibitor according to theinvention.

[0349] The dosage of the inhibitor compound of the invention will dependupon the nature and severity of the condition being treated, theadministration route and the size and species of the patient. However ingeneral, quantities of from 0.01 to 100 μmol/kg bodyweight will beadministered.

[0350] All publications referred to herein are hereby incorporated byreference.

[0351] The invention will now be described further with reference to thefollowing non-limiting Examples.

EXPERIMENTAL

[0352] Abbreviations used follow IUPAC-IUB nomencalture. The followingabbreviations are used throughout: aq. (aqueous), equiv, ([molar]equivalent), Boc (tertiary-butyloxycarbonyl), CMA (chloroform:methanol,concentrated ammonium hydroxide 80:18:2), DCC(1,3-dicyclohexylcarbodiimide), DCM (dichloromethane), DEPC (diethylcyanophosphonate), DIPEA (diisopropylethylamine), DMEA(dimethylethylamine), DMF (dimethylformamide), DMSO (dimethyl sulfoxide,perdeuterated if for NMR), EDCI(1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride), EtOAc(ethyl acetate), EtOH (ethanol), HATU([O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetra-methyluroniumhexafluorophosphate]), HOAt (1-hydroxy-7-aza-benzotriazole), HOBt(1-hydroxy-benzotriazole), HPLC (high-performance liquidchromatography), IS-MS (ion spray mass spectrum), RPHPLC (reverse phasehigh-performance liquid chromatography), SCX (strong cation exchangeresin), TFA (trifluoroacetic acid), THF (tetrahydrofuran), TLC (thinlayer chromatography with R_(f) as relative mobility), DSC (differentialscanning calorimetry), TGA (thermal gravimetric analysis).

[0353] All solution concentrations are expressed as % Vol./% Vol. unlessotherwise stated. Reagents were obtained from a variety of commercialsources.

[0354] IR means an infrared spectrum was obtained. ¹NMR, NMR, 1H-NMR, or1H NMR means a proton magnetic resonance spectrum was obtained.

[0355] HPLC Analysis (Methods A to D)

[0356] (Method A): Vydac C18 (4.6×250 mm), elute with a linear gradientof 90/10 through 50/50 (0.1% TFA in water/0.1% TFA in acetonitrile) over40 min, 1 mL/min.

[0357] (Method B): Waters Symmetry, C18 (4.6×250 mm) column. The elutionsystem consisted of linear gradient from 95:5 (0.2% TFA in H₂O)/(0.2%TFA in CH₃CN) to 5:95 (0.2% TFA in H₂O)/(0.2% TFA in CH₃CN) over 20 min,followed by (0.2% TFA in CH₃CN) isocratic over 15 min. The flow rate was1 ml/min. UV Detection was performed at 254 nm unless otherwise noted.

[0358] (Method C): Shimadzu LC6 gradient system equipped with anautosampler, a variable wavelength detector at flow rates of 0.4 ml/min.Eluant A consisted of aqTFA (0.1%) and eluant B 90% MeCN in aq TFA(0.1%) with gradient elution (0 min. 20% B then 20% to 100% over 15min.); Luna C18 (2.1×150 mm, 5 μM particle size).

[0359] (Method D): Microsorb-MV C18 (4.6×250 mm) column. The elutionsystem consisted of a linear gradient from 90:10 (2.5% TFA in H₂O):(2.5%TFA in acetonitrile) to 10:90 (2.5% TFA in H₂O):(2.5% TFA inacetonitrile) over 25 min at 30° C. and a flow rate of 1 mL/min. UVDetection was performed at 254 nm unless otherwise noted.

[0360] API-MS (atmospheric pressure chemical ionization mass spectra)were obtained on a PESciex API 150EX with a heated nebulizer andnitrogen as the reagent gas in positive ion mode.

[0361] CI-MS (Chemical ionization mass spectra) were obtained on aShimadzu 5000 direct insertion mass spectrometer in chemical ionizationmode utilizing methane as the reagent gas.

[0362] MALDI-TOF, Matrix assisted laser desorption ionisation—time offlight mass spectrometry, RT, retention time:

[0363] In general in this specification, “D-” or “R-” in the name of aproduct indicates the product was made beginning with a chiral startingmaterial, for example D-phenylglycine.

[0364] Preparation of Starting Materials and Intermediates

[0365] Intermediate substituted glycine compounds for starting materialsand intermediates, including those in which the amino group and/or thecarboxy group is protected, conveniently may be prepared using one ofthe procedures below, or by a similar procedure. It may be convenient orpreferred to change the order of steps in the preparation of a compoundof the invention and to use a similar procedure with a differentintermediate. In particular, it may be convenient to use an acyl groupR₂—CO— initially in a preparation, rather than an amino protectinggroup.

[0366] Abbreviations, in addition to others listed herein, include:TEMPO: 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical; (DHQD)₂PHAL:hydroquinidine 1,4-phthalazinediyl diether; r.b. or rb, round bottomed;PPh₃, triphenylphosphine; Boc₂O or Boc anhydride: di-tert-butyldicarbonate.

[0367] Preparation of Intermediates KE-1-KE-5

[0368] The following compounds were prepared according to the indicatedmethod (Method KE-A) from the indicated starting materials, unlessotherwise described.

[0369] Intermediate KE-1

[0370] Ethyl oxo-quinolin-8-ylacetate.

Method KE-A

[0371] To a stirring solution of 8-bromoquinoline (10.1 g, 48.5 mmol) inTHF (500 mL) at −78° C. was added dropwise a 1.3 M solution of sec-butyllithium (37.3 mL, 48.5 mmol) in cyclohexane. After 5 min, diethyloxalate (8 mL, 58.3 mmol) was added; and the solution was allowed toslowly warm to room temperature overnight. The next morning, thereaction was quenched with the addition of saturated aqueous NH₄Cl; andthe solvent was removed in vacuo. The residue was partitioned betweenethyl acetate and satd aq. NaHCO₃; the layers were separated; and thenthe aqueous phase was washed with brine, dried with MgSO₄, filtered andconcentrated in vacuo. The residue was chromatographed over silica gel,eluting with 20% ethyl acetate/hexanes through 25% ethylacetate/hexanes. The product containing fractions were combined andconcentrated in vacuo to give 5.88 g (53%) of the title compound.

[0372] 1H-NMR

[0373] IS-MS, m/e 230.1 (M+1)

[0374] Intermediate KE-2

[0375] Ethyl oxo-quinolin-5-ylacetate.

[0376] Prepared from 5-bromoquinoline and diethyl oxalate using MethodKE-A.

[0377] 1H-NMR

[0378] IS-MS, m/e 230.0 (M+1)

[0379] Intermediate KE-3

[0380] Ethyl oxo-thiazol-5-ylacetate.

[0381] To a r.b. flask (500 cm³) under argon, fitted with ethanolthermometer, septum cap, and dropping funnel, was added anhydrous ether(100 cm³) with stirring. This was cooled to −78° C. and 2 Mn-butyllithium (60 cm³, 120 mmol) was added.

[0382] A solution of silyl thiazole (16 g, 16 cm³, 100 mmol) inanhydrous ether (100 cm³) was then added by dropping funnel over 30minutes. This was allowed to stir for 1 hour to give a peach suspension.To this was added diethyl oxalate (16.3 cm³, 17.5 g, 120 mmol) rapidlyto give a brown solution, resulting in a temperature increase to −30° C.This was allowed to cool back to −78° C. and stirred for 30 minutes.Reaction monitored by ¹H NMR (CDCl₃).

[0383] The brown solution was poured onto 5% hydrochloric acid solution(300 cm³) with vigorous stirring for 30 minutes. Ether layer wasseparated and washed with saturated bicarbonate (ca. 80 cm³), dried overmagnesium sulphate, and concentrated in vacuo to give an orange oil.This was purified by flash chromatography (10% ethyl acetate/hexane) togive a yellow oil (7.31 g, 39.47 mmol) [40% Yield].

[0384]¹H NMR (CDCl₃); 1.42 (3H, t), 4.45 (2H, q), 8.89 (1H, s), 9.10(1H, s).

[0385] Intermediate KE-4

[0386] Ethyl oxo-thiazol-2-ylacetate.

[0387] Prepared from thiazole and diethyl oxalate using Method KE-A. Inthis case the temperature was held at −35° C. and n-butyllithium inhexane was used in place of sec-butyllithium in cyclohexane.

[0388]¹NMR

[0389] IS-MS, m/e 165.0 (M+1)

[0390] Intermediate KE-5

[0391] Ethyl oxo-isoquinolin-8-ylacetate.

[0392] Prepared from 8-bromoisoquinoline and diethyl oxalate usingMethod KE-A, substituting n-butyl lithium in hexanes for sec-butyllithium in cyclohexane.

[0393]¹NMR

[0394] IS-MS, m/e 230.0 (M+1)

[0395] Analysis for C₁₃H₁₁NO₃: Calcd: C, 68.11; H, 4.84; N, 6.11; Found:C, 68.11; H, 5.00; N, 6.14.

[0396] Preparation of Intermediates OX-1-OX-9

[0397] The following compounds were prepared according to the indicatedmethod (Method OX-A or Method OX-B) from the indicated startingmaterials unless otherwise described.

[0398] Intermediate OX-1

[0399] Ethyl Hydroxyimino-pyridin-2-ylacetate.

Method OX-A

[0400] To a stirring solution of ethyl 2-pyridylacetate (12.6 g, 76.3mmol) in acetic acid (19 mL) at 5° C. was added a solution of sodiumnitrite (6.05 g, 87.7 mmol) in water (12 mL) at a rate sufficient tomaintain the internal temperature below 15° C. After complete additionand an additional 30 min, an additional 30 mL of water were added. Theresulting white precipitate was filtered, washed with water, satd aq.NaHCO₃, and again with water. The solid was then dried under vacuum togive 14.1 g (95%) of the title compound.

[0401] 1H-NMR

[0402] IS-MS, m/e 194.9 (M+1)

[0403] Analysis for C₉H₁₀N₂O₃: Calcd: C, 55.67; H, 5.19; N, 14.43;Found: C, 55.79; H, 5.14; N, 14.13.

[0404] Intermediate OX-2

[0405] Ethyl Hydroxyimino-pyridin-3-ylacetate.

[0406] Using the procedure of Tikk et al [Acta. Chimica, Hungarica,114(3-4), 355], a mixture of ethyl hydroxyimino-pyridin-3-yl-acetate andn-butyl hydroxyimino-pyridin-3-yl-acetate was prepared from ethyl3-pyridinylacetate and n-butyl nitrite.

[0407] 1H-NMR

[0408] IS-MS, m/e 195 (M+1), 223.1 (M+1)

[0409] Intermediate OX-3

[0410] Ethyl Hydroxyimino-quinolin-8-ylacetate.

Method OX-B

[0411] To a stirring solution of ethyl oxo-quinolin-8-yl-acetate (5.5 g,24 mmol) in ethanol (140 mL) was added sodium acetate (2.16 g, 26.4mmol) followed by hydroxylamine hydrochloride (2.67 g, 38.4 mmol). Themixture was heated to reflux; and, after 7 h, the heating mantle wasremoved and the solution was allowed to stir overnight at roomtemperature. The next morning, the solvent was removed in vacuo and theresidue was partitioned between ethyl acetate and satd aq. NaHCO₃. Thelayers were separated and the organic phase was washed with brine, driedwith Na₂SO₄, filtered and concentrated in vacuo. The resulting foam wasrecrystalized from dichloromethane/hexanes to give an initial crop of2.5 g of the title compound as an off-white solid, followed by 0.31 g ofa second crop. The mother liquor was then concentrated in vacuo, theresidue was dissolved in a minimal amount of dichloromethane. Thesolution was then chromatographed over silica gel, eluting with 30%ethyl acetate/hexanes, then 40% ethyl acetate/hexanes, and finally withethyl acetate. The product containing fractions were combined andconcentrated in vacuo to give 1.94 g of the title compound for acombined yield of 4.75 g (81%).

[0412] 1H-NMR

[0413] IS-MS, m/e 245.0 (M+1)

[0414] Intermediate OX-4

[0415] Ethyl Hydroxyimino-quinolin-5-ylacetate.

[0416] Prepared from ethyl oxo-quinolin-5-yl-acetate using Method OX-B.

[0417] 1H-NMR

[0418] IS-MS, m/e 245.0 (M+1)

[0419] Intermediate OX-5

[0420] Ethyl Hydroxyimino-thiazol-5-ylacetate.

[0421] To a r.b. flask (500 cm³) was added the ethyloxo-thiazol-5-ylacetate (6.30 g, 34.02 mmol) to ethanol (ca. 180 cm³)with stirring. Sodium acetate (3.06 g, 37.30 mmol) and hydroxylaminehydrochloride (3.78 g, 54.43 mmol) were then added to give an off-whitesuspension. This was brought to reflux at 85° C. for 1 hour. Reactionmonitored by TLC (60% hexane/ethyl acetate; s.m. r.f 0.5, prod. r.f.0.3.). Reaction cooled and concentrated in vacuo. Product taken up inethyl acetate (c.a. 200 cm³) and washed with 5% hydrochloric acidsolution. Ethyl acetate layer was dried over magnesium sulphate andevaporated to dryness to give a cream solid (6.372 g, 31.825 mmol) [94%Yield].

[0422]¹H NMR (CDCl₃); 1.40 (3H, m), 4.40 (2H, m), 8.06 (⅓H, s), 8.78(⅓H, s), 8.95 (⅔H, s), 8.98 (⅔H,s).

[0423] Intermediate OX-6

[0424] Ethyl α-Oximino-thiazole-4-acetate.

[0425] To a 2 necked r.b. flask (100 cm³) with ethanol thermometer,concentrated sulphuric acid (25 cm³) was added and cooled to 0° C. withstirring. To this solution was added the ethylα-oximino-2-aminothiazole-4-acetate (5.00 g, 23.231 mmol). Water (10cm³) was then added and cooled to −10° C. A solution of sodium nitrite(1.683 g, 24.393 mmol) in water (5 cm³) was then added slowly over anhour keeping the temperature below −5° C.

[0426] To a separate r.b. flask (500 cm³), water (180 cm³) was added andcooled to 3° C. The reaction solution was poured in to the cold waterwith stirring and then cooled to −5° C. To this solution, 50%hypophosphoric acid (90 cm³) was added dropwise over 10 minutes keepingthe temperature at −5° C. The solution was allowed to warm to roomtemperature and stirred overnight. The product was extracted withdiethyl ether (ca. 3×150 cm³) and washed with water. The ether layer wasconcentrated in vacuo and treated to flash chromatography (50% ethylacetate/n-hexane) to yield a orange oil upon concentration in vacuo(0.60 g, 3.00 mmol) [13% yield]

[0427]¹H NMR (CDCl₃) 1.35 (3H, m), 4.35 (2H, m), 8.4 (1H, s), 8.9 (1H,s), 14.4 (1H, s).

[0428] Intermediate OX-7

[0429] Ethyl α-oximino-2-methylthiazole-4-acetate.

[0430] This was prepared from ethyl-γ-chloro-α-oximino-acetoacetate(1.44 g) using the method of Hatanaka et al. (Journal of MedicinalChemistry, 1973, 16(9), 978-984) to yield the titled compound (0.64 g).

[0431]¹H NMR (CDCl₃) 1.35 (3H, t), 2.7 (3H, s), 4.35 (2H, q), 8.2 (1H,s).

[0432] Ethyl γ-Chloro-α-oximinoacetoacetate.

[0433] This was prepared from ethyl oximinoacetoacetate (1.73 g) usingthe method of Hatanaka et al. (Journal of Medicinal Chemistry, 1973,16(9), 978-984) to yield the titled compound (1.44 g).

[0434]¹H NMR (CDCl₃) 1.25 (3H, t), 4.3 (2H, q), 4.55 (2H, s), 9.45 (1H,s), contains 20% starting material by NMR.

[0435] Ethyl Oximinoacetoacetate

[0436] This was prepared from ethyl acetoacetate (10.00 g) using themethod of Fischer (Organic Synthesis Coll. Vol. 3, 513-516) to yield thetitled compound (12.45 g).

[0437]¹H NMR (CDCl₃) 1.25 (3H, t), 2.35 (3H, S), 4.3 (2H, q), 8.8 (1H,br.).

[0438] Intermediate OX-8

[0439] Ethyl hydroxyimino-thiazol-2-ylacetate.

[0440] Prepared from ethyl oxo-thiazol-2-ylacetate using Method OX-B.

[0441]¹NMR

[0442] IS-MS, m/e 198.9(M−1)

[0443] Intermediate OX-9

[0444] Ethyl hydroxyimino-isoquinolin-8-ylacetate.

[0445] Prepared from ethyl oxo-isoquinolin-8-ylacetate using MethodOX-B.

[0446]¹NMR

[0447] IS-MS, m/e 245.0(M+1)

[0448] Analysis for C₁₃H₁₂N₂O₃: Calcd: C, 63.93; H, 4.95; N, 11.47;Found: C, 63.68; H, 4.60; N, 11.34.

[0449] Preparation of Intermediates AL-1-AL-3

[0450] The following compounds were prepared according to the indicatedmethod (Method AL-A or Method AL-B) from the indicated startingmaterials, unless otherwise described.

[0451] Intermediate AL-1

[0452] R-3-Bromo-(1-t-butoxycarbonylamino-2-hydroxyethyl)benzene.

Method AL-A

[0453] Sodium hydroxide (3.33 g, 83.25 mmol) was dissolved in water (220mL), and 20 mL of the resulting solution was removed and added topotassium osmate (410 mg, 1.11 mmol). The remaining sodium hydroxidesolution (200 mL) was added to a stirred solution of t-butyl carbamate(9.9 g, 84.5 mmol) in n-propanol (110 mL) followed by freshly preparedt-butyl hypochlorite (9.65 mL; 83.5 mmol). After stirring for 5 min, thesolution was cooled to 0° C. A solution of (DHQD)₂PHAL (1.30 g, 1.67mmol) in n-propanol (110 mL) was added, followed by a solution of3-bromostyrene (5 g, 27.31 mmol) in n-propanol (220 mL), followed bydropwise addition of the potassium osmate/sodium hydroxide solution. Thereaction was stirred overnight. Saturated aqueous sodium sulfite (150mL) was added, and the reaction was stirred for 15 min. The aqueouslayer was separated and extracted with ethyl acetate (3×200 mL). Thecombined organic layers were washed with brine and dried over MgSO₄.Removal of solvent under vacuum gave the crude product which waspurified by chromatography (silica, 3:2 hexane:ethyl acetate thenrechromatographed loading with toluene, gradient elution with hexane—4:1hexane:ethyl acetate) to give the title product (4.18 g, 49%).

[0454] Melting Point=90-91° C.

[0455]¹H NMR (CDCl₃).

[0456] Intermediate AL-2

[0457]R-3-Methoxycarbonyl-(1-t-butoxycarbonylamino-2-hydroxy-ethyl)benzene.

Method AL-B

[0458] In a glass liner containing a stirrer bar was placed Pd(OAc)₂(871 mg, 3.88 mmol), PPh₃ (1.96 g, 7.47 mmol, NaOAc (1.48 g, 18.04 mmol)and DMF (82 mL). To this stirred solution was added a solution ofR-3-bromo-(1-t-butoxy-carbonylamino-2-hydroxyethyl)benzene (4.27 g, 13.5mmol) in MeOH (82 mL). The resulting solution was purged with nitrogenand placed in a stirred pressure vessel. The system was charged to 4.1bar (60 psig) of CO and heated at 95° C. for 36 h. The mixture wascooled to room temperature, filtered through diatomaceous earth, andpartitioned between ethyl acetate and water. The organic layer waswashed with water (3×) and brine (1×) and dried over MgSO₄. Removal ofsolvent under vacuum gave the crude product which was purified bychromatography (silica gel, gradient elution with 30-35% ethylacetate/hexane) to provide the title product (3.53 g, 89%).

[0459] Melting Point=73-75° C. with decomposition

[0460]¹H NMR (CDCl₃).

[0461] API-MS, m/e=240 (M-C₄H₉+1).

[0462] Intermediate AL-3

[0463] R-3-Cyano-(1-t-butoxycarbonylamino-2-hydroxyethyl)benzene.

[0464] Prepared from 3-cyanostyrene using Method AL-A. 3-Cyanostyrenewas prepared using the method described below.

[0465] Melting Point=76° C.

[0466]¹H NMR (CDCl₃).

[0467] Preparation of 3-Cyanostyrene.

[0468] To a stirred suspension of methyltriphenylphosphonium bromide (75g, 209.71 mmol) in dry THF (750 mL) at 0° C. under nitrogen was addeddropwise n-BuLi (83 mL, 2.5 M in hexanes, 207.50 mmol). The mixture waswarmed to room temperature. 3-Cyanobenzaldehyde (25 g, 190.65 mmol) wasadded as a solid in 5 g batches, and the mixture was stirred at roomtemperature overnight. The reaction was quenched in water, and thesolvent was removed under vacuum. The residue was dissolved in theminimal amount of THF, and triphenylphosphine oxide was precipitatedusing ether. The solid was filtered through diatomaceous earth, and thefiltrate was concentrated. Distillation by Kugelrhor at 90° C./33 Pa(0.25 mm Hg) gave the product as a colorless oil (15.5 g, 62%).

[0469] Boiling Point=90° C. at 0.25 mmHg.

[0470]¹H NMR (CDCl₃).

[0471] Preparation of Intermediates PAE-1-PAE-18

[0472] The following compounds were prepared according to the indicatedmethod (Method PAE-A, Method PAE-B, Method PAE-C, Method PAE-D or PAE-E)from the indicated starting materials, unless otherwise described.

[0473] Intermediate PAE-1

[0474] Boc-D,L-(2-pyridinyl)glycine Ethyl Ester.

Method PAE-A

[0475] To a solution of ethyl hydroxyimino-pyridin-2-yl-acetate (7.8 g,40.15 g) in ethanol (175 mL) and glacial acetic acid (20 mL) was added5% Pd/C, and the mixture was shaken in a hydrogenation apparatus underan atmosphere of hydrogen at 4.1 bar (45 psig) for 4 h. The mixture wasfiltered through diatomaceous earth and concentrated in vacuo. Theresidue was dissolved in THF/H₂O (1/1, 240 mL) and treated withdi-tert-butyl dicarbonate (14.23 g, 65.2 mmol) and sodium bicarbonate(27.4 g, 326 mmol). After stirring at room temperature for 2 h, thesolution was concentrated in vacuo and the residue was partitionedbetween EtOAc and water. The organic phase was washed with brine, driedover magnesium sulfate, filtered and concentrated in vacuo. The crudematerial was purified via chromatography over silica gel, eluting with astepwise gradient of 10-20% ethyl acetate in dichloromethane to give8.11 g (72%) of the title compound as a yellow oil.

[0476] 1H-NMR

[0477] IS-MS, m/e 281.1 (M+1)

[0478] Intermediate PAE-2

[0479] Boc-D,L-(3-pyridinyl)glycine Ethyl Ester.

[0480] Prepared from ethyl hydroxyimino-pyridin-3-ylacetate using MethodPAE-A.

[0481] 1H-NMR

[0482] IS-MS, m/e 281.1 (M+1)

[0483] Intermediate PAE-3

[0484] Boc-D,L-(8-quinolinyl)glycine Ethyl Ester.

Method PAE-B

[0485] To a stirring solution of ethyl hydroxyimino-quinolin-8-ylacetate(2.4 g, 9.8 mmol) in 50% aq. formic acid (50 mL) at 0° C. was added zincdust (2 g, 31 mmol). After 1 min, the mixture was filtered throughdiatomaceous earth and the filtrate was loaded onto an SCX column. Afterwashing the column with methanol, the product was eluted with a 3 to 1mixture of dichloromethane and (2 N NH₃ in methanol). The productcontaining fractions were combined and concentrated in vacuo to give2.24 g of light orange oil (IS-MS, m/e 231.0 (M+1)).

[0486] The oil (2.14 g, 9.3 mmol) was dissolved in THF (40 mL) and tothis stirring solution was added triethylamine (1.4 mL, 10.2 mmol),followed by di-tert-butyl dicarbonate (2.1 g, 9.8 mmol). After 45 min,the solvent was removed in vacuo and the residue was partitioned betweenethyl acetate and water. The organic phase was then washed with satd aq.NaHCO₃, dried with Na₂SO₄, filtered and concentrated in vacuo. Theresidue was dissolved in a minimum volume of dichloromethane andchromatographed over silica gel, eluting with 5% ethyl acetate inhexanes. The product containing fractions were combined and concentratedto give 2.5 g (81%) of the title compound.

[0487] 1H-NMR

[0488] IS-MS, m/e 331.0 (M+1)

[0489] Intermediate PAE-4

[0490] Boc-D,L-(5-quinolinyl)glycine Ethyl Ester

[0491] Prepared from ethyl hydroxyimino-quinolin-5-ylacetate usingMethod PAE-B.

[0492] 1H-NMR

[0493] IS-MS, m/e 331.0 (M+1)

[0494] Intermediate PAE-5

[0495]N-4-Methoxybenzoyl-N-2,4-dimethoxybenzyl-D,L-(2-trifluoro-methylphenyl)glycineMethyl Ester.

Method PAE-C

[0496] To 2-trifluoromethylbenzaldehyde (1 g, 5.7 mmol) with stirringwas added 2,4-dimethoxybenzylamine (0.86 mL, 5.7 mmol) and methanol (2mL). After 5 min, the solution was diluted with toluene 100 mL andconcentrated in vacuo (twice). The residue was then dissolved inanhydrous methanol (12 mL) and 1,1-dimethyl-2-(methoxycarbonyloxy)ethylisonitrile [Tetrahedron, 55 (1999) 7411-7420] (0.9 g, 5.7 mmol) wasadded, followed by 4-methoxybenzoic acid (0.87 g, 5.7 mmol) Afterstirring for 72 h, the solvent was removed in vacuo and the residue waschromatographed over silica gel, eluting with a step gradient of 30%ethyl acetate in hexanes through 50% ethyl acetate in hexanes. Theproduct containing fractions were combined and concentrated in vacuo;and then the residue was dissolved in ethyl acetate, washed with satdaq. NaHCO₃, dried with Na₂SO₄, filtered and concentrated to give 1.76 g(48%) of thick oil (NMR, IS-MS, m/e 633.0 (M+1)). The oil (0.5 g, 0.79mmol) was then dissolved in toluene (5 mL) and concentrated in vacuo(twice) to give a white foam. The residue was then dissolved in THF (3mL) and potassium tert-butoxide (0.11 g, 0.95 mmol) was added. After 15min, 12 N HCl (0.079 mL, 0.95 mmol) was added and the solution wasallowed to stand overnight in the refrigerator. The next morning, thesolvent was removed and the residue was chromatographed over silica gel,eluting with 30% ethyl acetate in hexanes. The product containingfractions were combined and concentrated to give 0.32 g (79%) of thetitle compound.

[0497] 1H-NMR

[0498] IS-MS, m/e 518.0 (M+1)

[0499] Intermediate PAE-6

[0500] BOC-D,L-(5-thiazolyl)glycine ethyl ester.

[0501] To a r.b. flask (250 cm³), D,L-(5-thiazolyl)glycine ethyl ester(4.60 g, 24.7 mmol) was added to tetrahydrofuran (c.a. 100 cm³) withstirring to give a yellow solution. BOC anhydride (5.439 g, 24.948 mmol)and triethyl amine (3.79 cm³, 2.75 g, 27.17 mmol) were then added withstirring for 1 hour. Reaction monitored by TLC (60% hexane/ethylacetate; s.m. r.f 0.05, prod. r.f. 0.5.). The reaction concentrated invacuo and product taken up in ethyl acetate (c.a. 150 cm³), washed with5% hydrochloric acid solution (ca. 30 cm³), and saturated bicarbonate(ca. 30 cm³). Ethyl acetate layer was dried over magnesium sulphate andevaporated to dryness to give an orange oil (7.42 g, ˜24.70 mmol) [˜100%Yield].

[0502]¹H NMR (CDCl₃); 1.30 (3H, t), 1.48 (9H, s), 4.28 (2H, q), 5.68(1H, br.), 7.88 (1H, s), 8.78 (1H, s).

[0503] D,L-(5-Thiazolyl)glycine Ethyl Ester.

[0504] To a r.b. flask (250 cm³), was added 5-thiazolyl-oximinoaceticacid ethyl ester (6.37 g, 31.825 mmol) to ethanol (c.a. 80 cm³) withstirring. 50% Formic acid solution (50 cm³) was added with zinc dust(5.10 g, 81.83 mmol) and allowed to stir overnight. Reaction monitoredby TLC (60% hexane/ethyl acetate; s.m. r.f 0.3, prod. r.f. 0.05.).Reaction solution filtered over diatomaceous earth and filtrateconcentrated in vacuo. This was basified to pH 9 with anhydrouspotassium carbonate and product taken up in 3:1 chloroform/isopropanolsolution (c.a. 200 cm³). This was washed with saturated bicarbonate(c.a. 50 cm³), dried over magnesium sulphate and concentrated in vacuoto give a brown oil (4.60 g, 24.70 mmol) [78% Yield]

[0505] 1H NMR (CDCl₃); 1.25 (3H, t), 1.95 (2H, br.), 4.22 (2H, q), 4.85(1H, s), 7.80 (1H, s), 8.70 (1H, s).

[0506] Intermediate PAE-7

[0507] N-Boc-D,L-(4-thiazolyl)glycine ethyl ester

[0508] To a solution of D,L-(4-thiazolyl)glycine ethyl ester (0.460 g,2.470 mmol) in tetrahydrofuran (20 cm³), was added di-tert-butyldicarbonate (0.530 g, 2.470 mmol) and triethylamine (0.344 cm³, 2.470mmol). This was allowed to stir for 1 hour and the solution concentratedin vacuo. The oil was taken up in ethyl acetate (c.a. 50 cm³) washedwith 0.5% hydrochloric acid solution (c.a. 20 cm³), and saturated sodiumbicarbonate solution (c.a. 20 cm³). This was then dried over magnesiumsulphate and concentrated in vacuo to yield an orange oil (0.709 g,2.477 mmol) [˜100% yield].

[0509]¹H NMR (CDCl₃) 1.15 (3H, t), 1.35 (9H, s), 4.1 (2H, m), 5.45 (1H,d), 5.75 (1H, d), 7.3 (1H, d), 8.7 (1H, d).

[0510] D,L-(4-Thiazolyl)glycine Ethyl Ester.

[0511] This was prepared from ethyl-α-oximino-thiazole-4-acetate (0.60g) using the method of Hatanaka et al. (Journal of Medicinal Chemistry,1973, 16(9), 978-984) to yield the titled compound (0.46 g).

[0512]¹H NMR (CDCl₃) 1.25 (3H, t), 1.8-2.3 (2H, br.), 4.1 (2H, m), 4.75(1H, s), 7.25 (1H, d), 8.7 (1H, d).

[0513] Intermediate PAE-8

[0514] N-Boc-D,L-(2-methylthiazol-4-yl)glycine Ethyl Ester

[0515] To a solution of D,L-(2-methylthiazol-4-yl)glycine ethyl ester(0.397 g, 1.982 mmol) in tetrahydrofuran (20 cm³), was addeddi-tert-butyl dicarbonate (0.475 g, 2.180 mmol) and triethylamine (0.304cm³, 2.180 mmol). This was allowed to stir for 1 hour and the solutionconcentrated in vacuo. The oil was taken up in ethyl acetate (c.a. 50cm³) washed with 0.5% hydrochloric acid solution (c.a. 20 cm³), andsaturated sodium bicarbonate solution (c.a. 20 cm³). This was then driedover magnesium sulphate and concentrated in vacuo to yield a yellow oil(0.654 g, 2.177 mmol) [˜100% yield].

[0516]¹H NMR (CDCl₃) 1.1 (3H, s), 1.35 (9H, s), 2.6 (3H, s), 4.15 (3H,m), 5.3 (1H, d), 5.7 (1H, s), 7.0 (1H, s).

[0517] D,L-(2-Methylthiazol-4-yl)glycine Ethyl Ester.

[0518] This was prepared from ethyl-α-oximino-2-methylthiazole-4-acetate(0.62 g) using the method of Hatanaka et al. (Journal of MedicinalChemistry, 1973, 16(9), 978-984) to yield the titled compound (0.40 g).

[0519]¹H NMR (CDCl₃) 1.15 (3H, t), 1.95 (2H, br.), 2.6 (3H, s), 4.15(2H, m), 4.65 (1H, s), 6.95 (1H, s).

[0520] Intermediate PAE-9

[0521] Boc-R-(4-Hydroxyphenyl)glycine Methyl Ester

[0522] To a stirred mixture of R-(4-hydroxyphenyl)glycine methyl esterhydrochloride (14 g) and sodium bicarbonate (11.7 g) in THF (150 mL) andwater (50 mL), was added in one portion, di-t-butyl dicarbonate (15.9g). The mixture was stirred rapidly to allow thorough mixing for 4 h.Hexane (75 mL) was added and the organic layer separated and washed withsatd sodium bicarbonate solution, then brine and then dried withmagnesium sulphate. The drying agents was filtered off and washed with alittle THF and evaporated to dryness, finishing with a high vacuum pumpto remove the last traces of di-t-butyl dicarbonate. Yield 19.7 g, 96%.

[0523]¹H NMR

[0524] R-(4-Hydroxyphenyl) glycine Methyl Ester Hydrochloride.

[0525] To a dry 250 mL three necked round bottom flask, equipped with alow temperature thermometer, a septum for nitrogen coverage and anotherfor introduction of thionyl chloride by syringe, was addedR-4-hydroxyphenylglycine (12.5 g) and dry methanol (24 mL). The mixturewas stirred (magnetic stirrer) and cooled to an internal temperature of−20° C. using cardice/acetone. Using a syringe, thionyl chloride wasadded dropwise to the cooled mixture over a period of 10 min. (Care: thereaction of thionyl chloride with methanol is very exothermic and rateof addition should be such that the thionyl chloride is efficientlystirred into the mixture and that the temperature does not rise above−20° C. Once the addition was complete the mixture was allowed to warmto room temperature overnight (16-18 h). Dry ether (150 mL) was addedand the white ppt. that formed was filtered off, washed with a littlemore ether and dried. Yield 15.5 g, 95%.

[0526]¹H NMR

[0527] Intermediate PAE-10

[0528] Boc-R-(4-Trifluoromethanesulphonyloxyphenyl)glycine Methyl EsterHydrochloride.

[0529] To a stirred solution of Boc-R-(4-hydroxyphenyl)glycine methylester (19 g) in dichloromethane (400 mL) was added 2,6-lutidine (9.44mL) and 4-dimethylaminopyridine (1.65 g) and the mixture cooled in anice bath. Trifluoromethanane-sulphonic anhydride (13.74 mL) was addedover a period of 5 min, and then the reaction left to warm to roomtemperature over 4 h. The organic solution was washed with water (2×150mL), 1 N HCl (2×150 mL), and then saturated sodium bicarbonate (150 mL).The organics were dried with magnesium sulphate and then evaporated toan oil. The mixture was purified using flash chromatography (SiO₂ 250 g,eluting with 1:1 hexane/dichloromethane and then neat dichloromethane).Pure product fractions were combined and evaporated, finishing with ahigh vacuum pump to remove all traces of solvent, to give a white solid,19 g, 77%.

[0530]¹H NMR

[0531] Intermediate PAE-11

[0532] Boc-R-(4-Methoxycarbonylphenyl)glycine Methyl Ester.

Method PAE-D

[0533] Boc-R-4-trifluoromethanesulphonyloxyphenylglycine methyl ester(15 g), methanol (32.6 mL), bis-1,3-diphenyl-phosphinylpropane (448 mg),palladium (II) acetate (255 mg), triethylamine (10.2 mL) anddimethylformamide (72 mL) were placed in the glass liner of pressure(Parr) reactor and the reactor assembled. The vessel was pressurised to−0.68 bar (10 psig) with nitrogen and the gas released (repeated fivetimes to remove all oxygen from the system). Carbon monoxide gas wasthen carefully introduced (use extreme care—the gas cylinder ispressurised to far beyond the bursting disc pressure of the Parr,ideally use a pressure regulator to reduce the pressure to −6.8 bar, 100psig) to ˜1.4 bar (20 psig) and released three times (into the back of afume hood). Carbon monoxide was then added to −6.8 bar (100 psig) andthe stirrer started. The vessel was slowly heated to 65° C. internaltemperature and then stirred at 65° C. overnight. (At the early stagesmore carbon monoxide was added to maintain ˜6.8 bar, 100 psig.) A samplewas removed after 18 h and examined by tlc. When complete, the reactionwas cooled to ˜30° C., the gas released and the vessel flushed fivetimes with nitrogen as before. The reaction mixture was partitionedbetween ethyl acetate and water, and the organic layer washed with 1 Mhydrochloric acid and then saturated sodium bicarbonate. The solutionwas dried with MgSO₄ and evaporated. Flash chromatography of theresulting oil gave the product, pure by tlc, 10.6 g, 90%.

[0534]¹H NMR

[0535] Intermediate PAE-12

[0536] Boc-R-(4-Benzyloxycarbonylphenyl)glycine Methyl Ester

[0537] Prepared from Boc-R-4-trifluoromethanesulphonyloxy phenylglycinemethyl ester and benzyl alcohol using Method PAE-D.

[0538]¹H NMR

[0539] Intermediate PAE-13

[0540] Boc-R-(4-Carboxyphenyl)glycine Methyl Ester.

[0541] Boc-R-(4-benzyloxycarbonylphenyl)glycine methyl ester (500 mg)was dissolved in THF containing Pd/C 10% (100 mg) and hydrogenated at 1atm for 2 h. Removal of the catalyst by filtration and evaporation ofsolvent gave Boc-R-(4-carboxy-phenyl)glycine methyl ester (330 mg, 87%).

[0542]¹H NMR

[0543] Intermediate PAE-14

[0544] Boc-R-(4-carboxamidophenyl)glycine Methyl Ester.

Method PAE-E

[0545] To a solution of Boc-R-(4-carboxyphenyl)glycine methyl ester (3.5g) in DMF (30 mL) was added EDCI (2.60 g, 1.36 mmol) and HOBt (1.4 g,10.4 mmol), and the mixture stirred for 10 min before cooling in a icebath and bubbling in ammonia gas for 5 min. The mixture was stirred for2 h at room temperature and then diluted with ethyl acetate and washedwith water. The aqueous solution was extracted with a little ethylacetate and the combined organics washed with brine. The organicsolution was evaporated to an oil which was purified by flashchromatography (SiO₂—dichloromethane/ethyl acetate 0-25%) to giveBoc-R-(4-carboxamidophenyl)glycine methyl ester (1.7 g, 48%).

[0546]¹H NMR

[0547] Intermediate PAE-15

[0548] Boc-R-(4-methylcarboxamidophenyl)glycine Methyl Ester.

[0549] Prepared from Boc-R-(4-carboxyphenyl)glycine methyl ester andmethylamine using Method PAE-E.

[0550]¹H NMR

[0551] Intermediate PAE-16

[0552]N-4-Methoxybenzoyl-N-2,4-dimethoxybenzyl-D,L-(quinolin-4-yl)glycineMethyl Ester.

[0553] Prepared from quinoline-4-carboxaldehyde using Method PAE-C.

[0554]¹H NMR

[0555] Intermediate PAE-17

[0556] Ethyl Boc-D,L-thiazol-2-ylglycine.

[0557] Prepared from ethyl hydroxyimino-thiazol-2-ylacetate using MethodPAE-B. In this case, reaction with Zn/formic acid was conducted over 15min.

[0558]¹NMR

[0559] IS-MS, m/e 287.0 (M+1)

[0560] Intermediate PAE-18

[0561] Ethyl Boc-D,L-isoquinolin-8-ylglycine.

[0562] Prepared from ethyl hydroxyimino-isoquinolin-8-ylacetate usingMethod PAE-B. In this case, reaction with Zn/formic acid was conductedover 30 min, followed by concentration and partitioning of the residuebetween 3/1 chloroform/isopropanol and satd aq. NaHCO₃. The Bocprotection was carried out as previously described. Purification wasperformed using silica gel chromatography (Biotage Quad System) elutingwith 10% ethyl acetate in methylene chloride.

[0563]¹NMR

[0564] IS-MS, m/e 331.0 (M+1)

[0565] Analysis for C₁₈H₂₂N₂O₄: Calcd: C, 65.44; H, 6.71; N, 8.48;Found: C, 65.05; H, 6.67; N, 8.49.

[0566] Preparation of Intermediates PAA-1-PAA-28

[0567] The following compounds were prepared according to the indicatedmethod (Method PAA-A, Method PAA-B, Method PAA-C, Method PAA-D, MethodPAA-E or Method PAA-F) from the indicated starting materials, unlessotherwise described.

[0568] Intermediate PAA-1

[0569] Boc-D,L-(2-chlorophenyl)glycine.

Method PAA-A

[0570] 2-Chlorobenzaldehyde (20 mmol, 2.252 mL) and2,4-di-methoxybenzylamine (20 mmol, 3.004 mL) were added together andstirred for 2 hours. DCM (5 mL) was added and any water separated andremoved. tert-Butyl isonitrile (20 mmol, 2.262 mL) was added and stirredfor 10 min, followed by acetic acid (20 mmol, 1.145 mL). Stirring wascontinued for 3 days. The reaction mixture was then treated with TFA (30mL) and triethylsilane (5 mL). After 3 h the mixture was evaporated todryness, 6 M HCl (100 mL) added, and the whole refluxed overnight at130° C., stirring rapidly. The mixture was allowed to cool and extractedwith EtOAc (50 mL×2); the aqueous fraction was evaporated to dryness andtreated with 2 M NaOH solution. The mixture was extracted with EtOAc (50mL×2); excess boc anhydride (5.2 g) in dioxane (20 mL) was added to theaqueous fraction and stirred overnight. The mixture was extracted withdiethyl ether (100 mL×2), acidified to pH 1 (conc HCl) and extractedwith EtOAc (50 mL×2). The combined organic fractions were washed withwater and evaporated to dryness under high vacuum. The productBoc-2-chlorophenylglycine (4.252 g, 74.5*)

[0571]¹H NMR (CD₃CN/D₂O) 7.3 (4H, m); 5.5 (1H, s); 1.3 (9H, s). MS 286(M+1)

[0572] Intermediate PAA-1′

[0573] (R)-Benzyloxycarbonyl-(2-chlorophenyl)glycine.

[0574] Prepared from 2-chlorostyrene using the method of Sharpless et alJ.A.C.S. (1998) Vol120 No.6 1207-1217.

[0575] Intermediate PAA-1, Alternative Preparation

[0576] Boc-D,L-(2-chlorophenyl)glycine.

[0577] Prepared from 2-chlorobenzaldehyde using method PAA-F. In thiscase, the reaction temperature was not controlled upon addition of2-chlorobenzaldehyde and the reaction was allowed to stir for 2 h.Extraction of the intermediate aminonitrile was performed with ethylether in place of ethyl acetate and was further purified by addition ofHCl gas to the ethereal extracts followed by decantation of the motherliquor to isolate the semisolid hydrochloride salt. BOC protection ofthe amino acid was performed from 0° C. to room temperature over aperiod of one hour and the final extraction was performed with ethylacetate in place of ethyl ether.

[0578]¹H-NMR

[0579] IS-MS m/e 284 (M−1)

[0580] Intermediate PAA-2

[0581] Boc-D,L-(3-fluorophenyl)glycine.

[0582] Prepared from 3-fluorobenzaldehyde using Method PAA-A.

[0583]¹H NMR (CD₃CN/D₂O) 7.3 (1H, m), 7.1(3H, m); 5.2 (1H, s); 1.3 (9H,s). MS 270 (M+1)

[0584] Intermediate PAA-3

[0585] Boc-D,L-(4-fluorophenyl)glycine.

[0586] Prepared from 4-fluorobenzaldehyde using Method PAA-A.

[0587]¹H NMR (CD₃CN/D₂O) 7.3 (2H, m); 6.9 (2H, m), 5.0 (1H, s); 1.3 (9H,s). MS 270 (M+1)

[0588] Intermediate PAA-4

[0589] Boc-D,L-(2-methylphenyl)glycine.

[0590] Prepared from 2-methylbenzaldehyde using Method PAA-A.

[0591]¹H NMR (CD₃CN/D₂O) 7.3 (4H, m); 5.5 (1H, s); 2.5 (3H, s); 1.3 (9H,s). MS 266 (M+1)

[0592] Intermediate PAA-5

[0593] Boc-D,L-(3-thienyl)glycine.

[0594] Prepared from 3-thiophenecarboxaldehyde using Method PAA-A.

[0595]¹H NMR (CD₃CN/D₂O) 7.5 (2H, m); 7.1 (1H, d); 5.3 (1H, S); 1.3 (9H,s). MS 258 (M+1)

[0596] Intermediate PAA-6

[0597] Boc-D,L-(2-fluorophenyl)glycine.

[0598] Was obtained by treating D,L-2-fluorophenylglycine (Aldrich) withBoc anhydride (1.1 eq) and 2 M NaOH (1 eq) in ethanol. Aqueous work upas described above yielded the protected amino acid.

[0599]¹H NMR

[0600] Intermediate PAA-7

[0601] Boc-D,L-(2-methoxyphenyl)glycine.

[0602] Prepared from 2-methoxybenzaldehyde using Method PAA-A.

[0603]¹H NMR

[0604] Intermediate PAA-7, Alternative Preparation

[0605] Boc-D,L-(2-methoxyphenyl)glycine.

[0606] Prepared from 2-methoxybenzaldehyde using method PAA-F. In thiscase, the reaction was cooled to 0° C. before addition of2-methoxybenzaldehyde and was then allowed to stir at room temperatureovernight. Extraction of the intermediate aminonitrile was performedwith ethyl ether in place of ethyl acetate and was further purified byaddition of 1 M HCl in ethyl ether followed by filtration of thecrystalline hydrochloride salt. BOC protection of the amino acid wasperformed from 0° C. to room temperature over a period of three hours,and the final extraction was performed with dichloromethane in place ofethyl ether.

[0607]¹H-NMR

[0608] IS-MS m/e 280.1 (M−1)

[0609] Analysis for C₁₄H₁₉NO₅ Calcd: C, 59.78; H, 6.81; N, 4.98; Found:C, 59.68; H, 6.78; N, 4.95.

[0610] Intermediate PAA-8

[0611] Boc-D,L-(2-trifluoromethyl)phenylglycine.

[0612] Prepared from 2-trifluoromethylbenzaldehyde using Method PAA-A.

[0613]¹H NMR

[0614] Intermediate PAA-8, Alternative Preparation

[0615] Boc-D,L-(2-trifluoromethylphenyl)glycine.

[0616] Prepared from 2-trifluoromethylbenzaldehyde using method PAA-F.In this case, the reaction temperature was not controlled upon additionof 2-trifluoromethylbenzaldehyde and the reaction was allowed to stirfor 2 h. Extraction of the intermediate aminonitrile was performed withethyl ether in place of ethyl acetate and was further purified byaddition of HCl gas to the ethereal extracts followed by decantation ofthe mother liquor to isolate the semisolid hydrochloride salt. BOCprotection of the amino acid was performed from 0° C. to roomtemperature over a period of one hour and the final extraction wasperformed with ethyl acetate in place of ethyl ether.

[0617]¹H-NMR

[0618] IS-MS m/e 318 (M−1)

[0619] Intermediate PAA-9

[0620] Boc-D,L-(8-quinolinyl)glycine.

Method PAA-B

[0621] To a stirring solution of Boc-D,L-(8-quinolinyl)glycine ethylester (2.29 g, 6.93 mmol) in 1,4-dioxane (11 mL) was added a solution ofLiOH hydrate (0.32 g, 7.6 mmol) in water. After 2 h, the solvents wereremoved in vacuo and the residue was dissolved in water and washed withdiethyl ether. The aqueous phase was then acidified to pH 3 with solidcitric acid and extracted with ethyl acetate. The organic phase was thenwashed with brine, dried with Na₂SO₄, filtered and concentrated to give2.06 g (98%) of the title compound.

[0622]¹H-NMR

[0623] IS-MS, m/e 303.0 (M+1)

[0624] Intermediate PAA-10

[0625] Boc-D,L-(5-quinolinyl)glycine.

[0626] Prepared from Boc-D,L-(5-quinolinyl)glycine ethyl ester singMethod PAA-B.

[0627] 1H-NMR

[0628] IS-MS, m/e 303.0 (M+1)

[0629] Intermediate PAA-11

[0630] Boc-D-(3-bromophenyl)glycine.

[0631] Prepared fromR-3-bromo-(1-t-butoxycarbonylamino-2-hydroxyethyl)benzene using MethodPAA-C.

[0632] Melting Point=130-132° C. with decomposition

[0633]¹H NMR (CDCl₃)

[0634] API-MS, m/e=286 (M-CO₂H+1)

[0635] Intermediate PAA-12

[0636] Boc-D-(3-methoxycarbonylphenyl)glycine.

Method PAA-C

[0637] To a stirred solution ofR-3-methoxycarbonyl-(1-t-butoxy-carbonylamino-2-hydroxyethyl)benzene(338 mg, 1.14 mmol) in acetone (7.2 mL) was added 5% NaHCO₃ (3 mL). Thereaction mixture was cooled to 0° C. To the stirred suspension was addedKBr (14 mg, 0.12 mmol), TEMPO (181 mg, 1.16 mmol) and NaOCl dropwise(2.81 mL, 5.25%). After 1 h at 0° C., TEMPO (136 mg, 0.88 mmol) andNaOCl (1.09 mL; 5.25%) were added. The reaction was stirred for afurther 0.5 h at 0° C. and 5% NaHCO₃ (4.3 mL) was added. The reactionwas allowed to warm to room temperature overnight. Acetone was removedunder vacuum and the crude product was partitioned between ethyl acetateand water. The aqueous layer was washed with ethyl acetate (2×) andacidified to pH 5 with 10% citric acid and extracted with ethyl acetate(4×). The combined organic extracts were dried over MgSO₄. Removal ofsolvent under vacuum gave the product (305 mg, 86%).

[0638]¹H NMR (CDCl₃)

[0639] API-MS, m/e=254 (M-C₄H₉+1)

[0640] Intermediate PAA-13

[0641] Boc-D-(3-cyanophenyl)glycine.

[0642] Prepared fromR-3-cyano-(1-t-butoxycarbonylamino-2-hydroxyethyl)benzene using MethodPAA-C.

[0643]¹H NMR (CDCl₃)

[0644] API-MS, m/e=221 (M-C₄H₉+1)

[0645] Intermediate PAA-14

[0646] Boc-D-(3-ethanesulfonylaminophenyl)glycine.

[0647] To a stirring solution of 3-(ethanesulfonylamino-phenyl)glycine(20 g, 77.43 mmol) and sodium carbonate (8.2 g, 77.43 mmol) in 3:1THF:water (200 mL) at 0° C., was added di-tert-butyl dicarbonate (18.5g, 85.17 mmol). After stirring for 30 min, the cold bath was removed;and after an additional 30 min at room temperature the solvent wasremoved; and the residue was partitioned between ethyl acetate andwater. The aqueous layer was acidified to pH 2 with KHSO₄ and extractedtwice with ethyl acetate. The combined ethyl acetate extracts werewashed with water, dried with Na₂SO₄, filtered and concentrated in vacuoto give 17.51 g (63*) of a white solid.

[0648] 1H-NMR

[0649] IS-MS, m/e 357.0 (M−1)

[0650] Intermediate PAA-15

[0651] N-Boc-D,L-(5-thiazolyl)glycine.

[0652] To a r.b. flask (150 cm³), was added Boc-D,L-(5-thiazolyl)glycineethyl ester (7.00 g, 24.70 mmol) to ethanol (c.a. 100 cm³) withstirring. 2 M Sodium hydroxide solution (25 cm³, 50 mmol) was added andallowed to stir for 1 h. Reaction monitored by TLC (60% hexane/ethylacetate; s.m. r.f 0.5, prod. r.f. 0.). Reaction concentrated in vacuoand product taken up in saturated bicarbonate (c.a. 50 cm³) and washedwith ethyl acetate (c.a. 30 cm³). Aqueous layer was acidified to pH 2with concentrated hydrochloric acid and product extracted with 3:1chloroform/isopropanol solution (c.a. 3×60 cm³). The organic layer wasdried over magnesium sulphate and evaporated to dryness to give anorange solid (4.47 g, 17.30 mmol) [74% Yield].

[0653]¹H NMR (CDCl₃); 1.35 (9H, s), 5.60 (1H, d), 5.83 (1H, d), 7.88(1H, s), 8.80 (1H, s)

[0654] Intermediate PAA-16

[0655] N-Boc-D,L-(4-thiazolyl)glycine.

Method PAA-D

[0656] To a solution of N-Boc-D,L-(4-thiazolyl)glycine ethyl ester(0.700 g, 2.470 mmol) in methanol (c.a. 15 cm³), was added 2 M sodiumhydroxide (2.47 cm³, 4.940 mmol) and allowed to stir for 90 min. Thesolution was concentrated in vacuo and taken up in water (c.a. 20 cm³).The aqueous solution was washed with ethyl acetate (c.a. 20 cm³), andthen acidified to pH 2 with 5% hydrochloric acid solution (c.a. 50 cm³).The product was extracted with ethyl acetate (c.a. 3×30 cm³), dried overmagnesium sulphate, and concentrated in vacuo to yield a pale yellow oil(0.582 g, 2.254 mmol) [91% yield]

[0657]¹H NMR (CDCl₃) 1.35 (9H, s), 5.5 (1H, d), 5.8 (1H, d), 7.35 (1H,d), 8.75 (1H, d), 9.8-10.2 (1H, br.)

[0658] Intermediate PAA-17

[0659] N-Boc-D,L-(2-methylthiazol-4-yl)glycine.

[0660] Prepared from N-Boc-D,L-(2-methylthiazol-4-yl)glycine ethyl esterusing Method PAA-D.

[0661]¹H NMR (CDCl₃) 1.35 (9H, s), 2.6 (3H, s), 5.4 (1H, d), 5.9 (1H,s), 7.1 (1H, s).

[0662] Intermediate PAA-18

[0663] N-Boc-D,L-(2-Benzyloxycarbonylamino-4-thiazolyl)glycine.

[0664] Is prepared fromD,L-(2-benzyloxycarbonylamino-4-thiazolyl)glycine. The benzyloxycarbonylprotecting group is removed from the thiazolyl amino group at aconvenient point in the preparation of a final compound using aconventional method, such as, for example, heating a solution of anintermediate in HBr/acetic acid at 60° C., followed by evaporation and aconventional isolation, such as by using SCX ion exchangechromatography.

[0665] D,L-(2-Benzyloxycarbonylamino-4-thiazolyl)glycine.

[0666] Was prepared by the method of Hardy, K.; Harrington, F. andStachulski, A.-J. Chem. Soc. Perkin Trans I (1984) 1227-1235.

[0667] Intermediate PAA-19

[0668] Boc-R-(4-methoxycarbonylphenyl)glycine.

[0669] To a solution of Boc-R-(4-methoxycarbonylphenyl)glycine methylester (692 mg) in THF (10 mL) was added a solution of lithium hydroxidehydrate (90 mg) in water (7 mL). The mixture immediately became cloudyand over 15 min cleared. After 30 min, tlc showed the reaction to becomplete. Ethyl acetate (20 mL) and water (20 mL) were added, and theaqueous layer separated. The aqueous solution was acidified with 2 Mhydrochloric acid and extracted with ethyl acetate (3×20 mL). Theorganic solution was then washed with water×2 and brine×2, dried withMgSO₄ and evaporated to give the mono-ester (650 mg, 98%), pure by tlc.

[0670]¹H NMR

[0671] Intermediate PAA-20

[0672] Boc-R-(4-Methoxyphenyl)glycine.

[0673] Boc-R-(4-hydroxyphenyl)glycine methyl ester was converted toBoc-R-4-methoxyphenylglycine using the alkylation method described byBasak et al.(Tetrahedron Lett. 1998, 39 (27), 4883-4886), followed byhydrolysis of the methyl ester with lithium hydroxide in aqueous THF.

[0674]¹H NMR

[0675] Intermediate PAA-21

[0676]N-4-Methoxybenzoyl-N-2,4-dimethoxybenzyl-D,L-(2-trifluoro-methylphenyl)glycine.

[0677] Prepared fromN-4-methoxybenzoyl-N-2,4-dimethoxybenzyl-D,L-(2-trifluoromethylphenyl)glycinemethyl ester using Method PAA-B (3 equivalents of LiOH hydrate).

[0678]¹H NMR

[0679] IS-MS, m/e 503.9 (m+1)

[0680] Intermediate PAA-22

[0681]N-4-Methoxybenzoyl-N-2,4-dimethoxybenzyl-D,L-(thien-2-yl)-glycine.

Method PAA-E

[0682] To a solution of 2-thiopheneboronic acid (5.0 g, 39.0 mmol, 1equiv) in 275 mL of methylene chloride at rt was added3,4-dimethoxybenzylamine (5.89 mL, 39.0 mmol, 1 equiv) followed byglyoxylic acid monohydrate 3.6 g, 39 mmol, 1 equiv). The reaction wasallowed to stir for 56 hours at rt after which time the resultantprecipitate was filtered and washed with methylene chloride to afford9.3 g (78%) of N-2,4-dimethoxybenzyl-D,L-(thien-2-yl)glycine as anoff-white solid (IS-MS, m/e 308 (m+1)).

[0683] A portion of the solid (5.0 g, 16.3 mmol, 1 equiv.) was dissolvedin acetone (20 mL) and 1 N sodium hydroxide (20 mL) at rt. To thissolution was simultaneously added anisoyl chloride (2.78 g, 16.3 mmol, 1equiv.) in 20 mL of acetone and 2 N sodium hydroxide in dropwisefashion. After stirring at rt for 1 h, the reaction was cooled to 0° C.and was acidified to pH 2-3. Diethyl ether was added and the product wasextracted into the organic phase. The combined organic phases werewashed with brine, dried over sodium sulfate, filtered, and concentratedto afford 5.1 g (71%) of the titled compound as a white solid.

[0684] IS-MS, m/e 440 (m+1).

[0685] Intermediate PAA-23

[0686] N-Boc-N-2,4-dimethoxybenzyl-D,L-(thien-2-yl)glycine.

[0687] To a solution of N-2,4-dimethoxybenzyl-D,L-(thien-2-yl)glycine(1.0 g, 3.2 mmol, 1 equiv) in 6 mL of acetone and 6 mL of water at rtwas added triethylamine (0.97 mL, 7.0 mmol, 2.1 equiv.) followed byaddition of 2-(tert-butoxy-carbonyloxyimino)-2-phenylacetonitrile(BOC-ON) (0.76 g, 3.1 mmol, 0.95 equiv). After stirring at rt overnight,the reaction was diluted with water and washed with ether. The aqueousphase was then acidified with 0.5 M citric acid and the product wasextracted into diethyl ether. The combined organic phases were washedwith brine, dried over sodium sulfate, filtered, and concentrated toafford 0.38 g (29%) of the titled compound as a crude yellow oil.

[0688] IS-MS, m/e 408 (m+1).

[0689] Intermediate PAA-24

[0690] Boc-D,L-isoquinolin-8-ylglycine.

[0691] Prepared from ethyl Boc-D,L-isoquinolin-8-ylglycine using MethodPAA-B. The product was precipitated from a basic aqueous solution byadjusting the pH to 3 with solid citric acid.

[0692]¹NMR

[0693] IS-MS, m/e 303.0 (M+1)

[0694] Analysis for C₁₆H₁₈N₂O₄.0.5H₂O: Calcd: C, 61.73; H, 6.15; N,9.00; Found: C, 61.62; H, 5.66; N, 8.84.

[0695] Intermediate PAA-25

[0696] Boc-D,L-Naphthalen-1-ylglycine.

Method PAA-F

[0697] Part A: D,L-Naphthalen-1-ylglycine hydrochloride.

[0698] To a solution of sodium cyanide (10.0 g, 0.22 mmol) in 40 mL ofwater was added ammonium chloride (11.4 g, 0.22 mmol), and the mixturewas stirred until dissolution was complete. A solution of1-naphthaldehyde (31.0 g, 0.22 mmol) in 40 mL of methanol was then addedand the resultant mixture was allowed to stir at room temperature fortwo days. An additional 150 mL of water was then added and the crudeproduct was extracted into EtOAc. The combined organic layers werewashed with water, dried over Na₂SO₄, filtered and concentrated toafford a crude oil. The crude residue was chromatographed over silicagel, eluting with with 10:1 EtOAc:CH₂Cl₂, to give 35 g of a light brownoil. This material was then dissolved in 250 mL of 5 N HCl and washeated to reflux for 9 h. The reaction was allowed to cool to roomtemperature and the product was allowed to crystallize overnight.Filtration of the mixture afforded 13.6 g (29%) of the title compound aslight brown crystals.

[0699]¹NMR

[0700] IS-MS, m/e 201.9 (M+1)

[0701] Part B: Boc-D,L-Naphthalen-1-ylglycine.

[0702] To a solution of D,L-naphthalen-1-ylglycine hydrochloride (13.6g, 57.2 mmol) and 2 N sodium hydroxide (57 mL, 115 mmol) in 120 mL of1,4-dioxane and 60 mL of water was added (Boc)₂O (15 g, 69 mmol). Thereaction was allowed to stir at room temperature for 3 h after whichtime the solution was brought to pH 5 by addition of 1 N sulfuric acid.The product was then extracted into EtOAc; and the combined organicextracts were dried over Na₂SO₄, filtered, and concentrated to give 14 g(81%) of the title compound as a light brown foam.

[0703]¹NMR

[0704] IS-MS, m/e 300.1 (M−1)

[0705] Intermediate PAA-26

[0706] Boc-D,L-(2-methylthiophenyl)glycine.

[0707] To a solution of 2-(methylthio)benzaldehyde (15 g, 98.7 mmol) in100 mL of ethanol was added ammonium carbonate (23.1 g, 296 mmol) and asolution of potassium cyanide (12 g, 148 mmol) in 100 mL water. Thereaction was heated and stirred at 70° C. for 3 h after which time thereaction was concentrated under reduced pressure. The product wasextracted into ethyl acetate; and the combined organic phases werewashed with brine, dried over Na₂SO₄, filtered and concentrated. Theresultant crude residue was taken up in 70 mL of ethyl acetate, and 70mL of 5 N sodium hydroxide was added. The reaction was heated to refluxfor three days after which time the ethyl acetate was removed underreduced pressure. To the aqueous mixture was sequentially added 100 mLof dioxane, Boc₂O (42 g, 192 mmol), and 100 mL of 2.5 N sodiumhydroxide. The reaction was then heated at reflux-for 48 h. Aftercooling to room temperature, the reaction was diluted with water and theaqueous phase was washed with ethyl ether. The aqueous layer was thenacidified to pH 2 and the product was extracted into ethyl acetate. Thecombined organic extracts were washed with brine, dried over MgSO₄,filtered, and concentrated to afford 21.7 g of a crude residue.Purification by silica gel chromatography (gradient elution, 97:2:1 to95:4:1 dichloromethane:methanol:acetic acid) provided 5.0 g (17%) of thetitle compound.

[0708]¹H-NMR

[0709] ES-MS m/e 296 (M−1)

[0710] Intermediate PAA-27

[0711] Boc-D,L-(2-methylsulfonylphenyl)glycine.

[0712] To a solution of boc-D,L-(2-methylthiophenyl)glycine (4.5 g, 15.2mmol) in 75 mL of methanol was added a solution of oxone (14 g, 23 mmol)in water. The reaction was stirred at room temperature for 2 h afterwhich time the methanol was removed under reduced pressure. The productwas extracted into ethyl acetate and the combined organic layers werewashed with brine, dried over MgSO₄, filtered, and concentrated toafford 4.35 g (87%) of the title compound.

[0713]¹H-NMR

[0714] ES-MS m/e 230(M+1-C₅H₉O₂)

[0715] Intermediate PAA-28

[0716] Boc-D,L-(benzo[b]thiophen-3-yl)glycine.

[0717] May be prepared by the method of Kukolja, S. et al. J. Med. Chem.1985, 28, 1886-1896.

Preparation of Intermediates A-1-A-12

[0718] The following compounds were prepared according to the indicatedmethod (Method A-A or Method A-B) from the indicated starting materials,unless otherwise described.

[0719] Intermediate A-1

[0720] 1-[2-(4-Pyridinyl)ethyl]piperazine hydrochloride.

Method A-A

[0721] A. 1-Boc-piperazine (30 g, 285 mmol), 4-vinylpyridine (40 g, 216mmol) and acetic acid (12.9 g, 215 mmol) were mixed in ethanol (400 mL)and heated to reflux for 18 h. The mixture was cooled to roomtemperature and concentrated under vacuum. The residue was dissolved inwater and ethyl acetate and neutralized with satd NaHCO₃. The layerswere separated. The water layer was extracted with ethyl acetate. Theorganic layers were combined, washed with brine, dried (Na₂SO₄),filtered and concentrated. The crude product was purified by SiO₂chromatography to provide 1-Boc-4-[2-(4-pyridinyl)ethyl]piperazine (55.9g, 87%) as an off white solid.

[0722] 1H-NMR(CDCl₃)

[0723] CI-MS, m/e=292 (M+1)

[0724] B. 1-Boc-4-[2-(4-pyridinyl)ethyl]piperazine (25 g, 85.8 mmol) wasdissolved in methanol (100 mL) and was cooled to 0° C. Saturated HCl inmethanol (100 mL) was added, and the mixture allowed to warm to roomtemperature for 1 h. The mixture was concentrated under vacuum andprovided 1-[2-(4-pyridinyl)ethyl]piperazine hydrochloride (23.8 g, 92%)as a white solid.

[0725] 1H-NMR (CD₃OD)

[0726] CI-MS, m/e=192 (M+1)

[0727] Alternatively, 1-Boc-4-[2-(4-pyridinyl)ethyl]piperazine (1.0 g,3.43 mmol) was dissolved in ethyl ether. Ethyl acetate (15 mL) saturatedwith HCl was added, and the mixture stirred for 30 min at roomtemperature. The mixture was concentrated under vacuum and provided1-[2-(4-pyridinyl)-ethyl]piperazine hydrochloride (900 mg, 87%) as a tansolid.

[0728] 1H-NMR (CD₃OD)

[0729] CI-MS, m/e=192 (M+1)

[0730] Intermediate A-2

[0731] 1-[2-(2-Pyridinyl)ethyl]piperazine.

[0732] Prepared from Boc-piperazine and 2-vinylpyridine using MethodA-A.

[0733] 1H-NMR(CD₃OD)

[0734] CI-MS, m/e=192 (M+1)

[0735] Intermediate A-3

[0736] 1-[2-(2-Pyrazinyl)ethyl]piperazine.

[0737] Prepared from Boc-piperazine and 2-vinylpyrazine using MethodA-A.

[0738] 1H-NMR (CD₃OD)

[0739] CI-MS, m/e=193 (M+1)

[0740] Intermediate A-4

[0741] 1-[2-(3-Pyridazinyl)ethyl]piperazine.

[0742] Prepared from Boc-piperazine and 3-vinylpyridazine (preparedusing the method described in J. Chem. Soc., Chem. Commun. 1985,1632-1633) using Method A-A.

[0743] 1H NMR (CD₃OD)

[0744] API-MS, m/e=193 (M+1)

[0745] Intermediate A-5

[0746] 1-[2-(3-Pyridinyl)ethyl]piperazine.

Method A-B

[0747] 1-Boc-4-[(3-pyridinyl)acetyl]piperazine (8.0 g, 26.2 mmol) wasadded to a solution of borane-THF (2.0 M in THF, 39.5 mL, 78.6 mmol) inTHF (200 mL) at 0° C. The mixture was heated to reflux for 8 h andcooled to room temperature. The excess borane was quenched with methanoland 3 N HCl. The mixture stirred for 3 h at room temperature, and thesolvents were removed under vacuum. The crude product was purified bychromatography (SiO₂, 4:1 CH₂Cl₂:CMA) to provide1-[2-(3-pyridinyl)ethyl]piperazine (2.82 g, 36%) as a light yellow oil.

[0748] 1H NMR (CD₃OD)

[0749] API-MS, m/e=192 (M+1)

[0750] Intermediate A-6

[0751] 1-[2-(4-Imidazolyl)ethyl]piperazine.

[0752] Prepared from 1-Boc-4-[(4-imidazolyl)acetyl]piperazine usingMethod A-B.

[0753] 1H-NMR

[0754] IS-MS, m/e 181.2 (M+1)

[0755] Intermediate A-7

[0756] 1-[2-(1-Imidazolyl)ethyl]piperazine.

[0757] Prepared from 1-Boc-4-[(1-imidazolyl)acetyl]piperazine usingMethod A-B.

[0758] 1H-NMR

[0759] IS-MS, m/e 181.4 (M+1)

[0760] Intermediate A-8

[0761] 1-[2-(1-Pyrazolyl)ethyl]piperazine.

[0762] Prepared from 1-Boc-4-[(1-pyrazolyl)acetylpiperazine using MethodA-B.

[0763] 1H-NMR

[0764] IS-MS, m/e 181.4 (M+1)

[0765] Intermediate A-9

[0766] 1-(2-Thiazol-2-ylethyl)piperazine.

[0767] A. A solution of Boc₂O (26 g, 120 mmol) in methylene chloride (50mL) was slowly added to a solution of ethyl piperazin-1-ylacetate (20 g,116 mmol) in CH₂Cl₂ (500 mL). The mixture was stirred for 1 h at roomtemperature. The solvent was removed under vacuum to provide ethyl4-Boc-piperazin-1-ylacetate (31.9 g, 100%) as a white solid.

[0768]¹H NMR (CDCl₃).

[0769] CI-MS, m/e=273 (M+1).

[0770] B. N-Methoxy-N-methylamine hydrochloride (4.3 g, 44.1 mmol) wasdissolved in methylene chloride (25 mL). The solution was cooled to −78°C., and a solution of trimethylaluminum (44.1 mL, 44.1 mmol, 1 M inheptane) was slowly added. The mixture was stirred at 0° C. for 30 minand was allowed to warm to room temperature and stirred for 30 min. Themixture was then cooled to 0° C., and a solution of ethyl4-Boc-piperazin-1-ylacetate (10 g, 36.8 mmol) was added dropwise. After15 min, the cold bath was removed and stirring continued overnight. Themixture was diluted with ethyl acetate and water. The layers wereseparated, and the water layer extracted with ethyl acetate. The organiclayers were combined, washed with brine, dried (Na₂SO₄), filtered, andconcentrated to provide N-methoxy-N-methyl-4-Boc-piperazin-1-ylacetamideas a light yellow oil (6.02 g, 57%) that solidified upon standing. Theproduct was used without further purification.

[0771]¹H NMR (CDCl₃).

[0772] CI-MS, m/e=288 (M+1).

[0773] C. n-Butyl lithium (1 M in hexanes, 12.2 mL, 12.2 mmol) wasslowly added to a solution of 2-bromothiazole (2.0 g, 12.2 mmol) indiethyl ether (50 mL) at −78° C. The mixture stirred at −78° C. for 1 h.Then, a solution of N-methoxy-N-methyl-4-Boc-piperazin-1-ylacetamide(3.0 g, 10.4 mmol) in tetrahydrofuran was slowly added. The mixture wasallowed to slowly warm to −20° C. and stirred for 4 h. The mixture wasthen diluted with water followed by ethyl acetate. The water layer wasextracted with ethyl acetate and the organic layers were combined,washed with brine, dried (Na₂SO₄), filtered, and concentrated. The crudeproduct was purified by chromatography (SiO₂, 20:1-6:1 CH₂Cl₂:CMA) toprovide 1-Boc-4-(2-oxo-2-thiazol-2-ylethyl)piperazine (2.2 g, 68%) as acolorless oil.

[0774]¹H NMR (CDCl₃).

[0775] CI-MS, m/e 312 (M+1).

[0776] D. 1-Boc-4-(2-oxo-2-thiazol-2-ylethyl)piperazine (5.0 g, 16.1mmol) was dissolved in methanol (25 mL). To this solution, magnesiumsulfate (2 g) was added, followed by p-tosylhydrazine (3.9 g, 20.2mmol). The mixture was stirred for 48 h and then filtered, and thefiltrate concentrated under vacuum. The residue (6.0 g, about 12 mmol)was redissolved in methanol (120 mL), and sodium triacetoxyborohydride(10.1 g, 48 mmol) was added. The mixture was heated to reflux overnight.The mixture was cooled to 0° C. and concentrated HCl (15 mL) slowlyadded. The mixture was allowed to warm to room temperature and stirredfor 1 h. The mixture was concentrated to half the volume and placed onan SCX column (30 g, pretreated with 5% acetic acid in methanol) andwashed with methanol (500 mL). The product was eluted with saturatedammonium hydroxide in methanol (500 mL) and the solvent removed undervacuum. The crude product was then purified by chromatography (SiO₂,12:1-4:1 CH₂Cl₂:CMA) to provide 1-(2-thiazol-2-ylethyl)piperazine (1.9g, 57%).

[0777]¹H NMR (CDCl₃).

[0778] CI-MS, m/e=198 (M+1).

[0779] Intermediate A-10

[0780] 1-[2-(2-Benzyloxycarbonylaminothiazol-4-yl)ethyl]piperazineHydrochloride.

[0781] Using methods substantially equivalent to those described inMethod A-B, the title compound was prepared from1-Boc-4-[2-(2-benzyloxycarbonylaminothiazol-4-yl)acetyl]piperazine(85%).

[0782]¹H NMR (CD₃OD).

[0783] CI-MS, m/e=347 (M+1).

[0784] Intermediate A-11

[0785] 1-[2-(3-Fluoropyridin-4-yl)ethyl]piperazine Trihydrochloride.

[0786] A. n-Butyl lithium (1.8 M in hexanes, 35 mL, 64.4 mmol) wascharged to a round bottom flask and diluted with THF (25 mL). Thesolution was cooled to 0° C., and a solution of N,N-diisopropylamine(9.0 mL, 65 mmol) in THF (25 mL) was slowly added. The mixture wasstirred at 0° C. for 20 min and was cooled to −78° C. A solution of3-fluoropyridine (20 g, 206 mmol) was added dropwise (temperature ofmixture kept below −70° C.), resulting in the formation of a redprecipitate. The mixture was stirred at −78° C. for 4 h. Ethylene oxide(4.6 M in THF, 67.2 mL, 309 mmol) was slowly added, and the mixtureallowed to warm to room temperature overnight. The mixture was dilutedwith water and CH₂Cl₂. The layers were separated, and the water layerextracted with CH₂Cl₂. The organic layers were combined, washed withbrine, dried (Na₂SO₄), filtered, and concentrated to provide a darkbrown oil. The residue was purified by chromatography (SiO₂, 19:1-6:1CH₂Cl₂:CMA) to provide 3-fluoro-4-(2-hydroxyethyl)pyridine (6.7 g, 23%)as a tan oil.

[0787]¹H NMR (CDCl₃).

[0788] CI-MS, m/e=142 (M+1).

[0789] B. 3-Fluoro-4-(2-hydroxyethyl)pyridine (4.0 g, 28.3 mmol) andtriethylamine (8.3 mL, 60 mmol) were dissolved in CH₂Cl₂ (40 mL) andcooled to 0° C. To this solution, methanesulfonyl chloride (2.0 mL, 31.2mmol) was added dropwise. The mixture stirred at 0° C. for 1 h. Themixture was diluted with water, and the layers separated. The waterlayer was extracted with CH₂Cl₂. The organic layers were combined,washed with brine, dried (Na₂SO₄), filtered, and concentrated undervacuum to provide 3-fluoro-4-(2-methanesulfonyloxyethyl)pyridine (5.2 g,83%) as a pink oil.

[0790]¹H NMR (CDCl₃).

[0791] CI-MS, m/e=220 (M+1).

[0792] C. 3-Fluoro-4-(2-methanesulfonyloxyethyl)pyridine (5.2 g, 23.7mmol) was dissolved in DMF (65 mL). 1-Boc-piperazine (8.85 g, 47.4mmol), K₂CO₃ (3.3 g, 23.7 mmol), NaI (3.6 g, 23.7 mmol) and Cs₂CO₃ (7.7g, 23.7 mmol) were added, and the mixture heated to 55° C. for 18 h. Themixture was cooled to room temperature and diluted with water and ethylacetate. The layers were separated, and the water layer extracted withethyl acetate. The organic layers were combined, washed with brine,dried (Na₂SO₄), filtered, and concentrated under vacuum to provide asolution of product and DMF. The residue was dissolved in diethyl etherand water. The layers were separated, and the water layer extracted withdiethyl ether. The organic layers were combined, washed with brine,dried (Na₂SO₄), filtered, and concentrated under vacuum. The crudeproduct was purified by chromatography (SiO₂, 15:1-6:1 CH₂Cl₂:CMA) togive 1-Boc-4-[2-(3-fluoropyridin-4-yl)ethyl]piperazine (6.2 g, 84%).

[0793]¹H NMR (CDCl₃).

[0794] CI-MS, m/e=310 (M+1).

[0795] D. 1-Boc-4-[2-(3-fluoropyridin-4-yl)ethyl]piperazine (6.0 g, 19.4mmol) was dissolved in methanol (20 mL) and anisole (6 mL). To thissolution, concentrated hydrochloric acid (15 mL) was added and themixture stirred for 1 h. The solvents were removed under vacuum toprovide a yellow solid. The residue was suspended in diethyl ether andsonicated for 1 h. The product was recovered by vacuum filtration, andthe solid dried under vacuum to provide1-[2-(3-fluoropyridin-4-yl)ethyl]piperazine trihydrochloride (5.2 g,84%) as an off white solid.

[0796]¹H NMR (CD₃OD).

[0797] CI-MS, m/e=210 (C₁₁H₁₆FN₃+1).

[0798] Intermediate A-12

[0799] 1-[2-(2-Cyanopyridin-4-yl)ethyl]piperazine.

[0800] A. Ethyl 4-pyridylacetate (20 g, 121 mmol) was added to asuspension of LiAlH₄ (9.2 g, 242 mmol) in diethyl ether (600 mL).Diatomaceous earth (about 50 mL) was added to aid stirring. The mixturestirred overnight. The mixture was cooled to 0° C. and aqueous NaOH(15%) was added until degassing ceased. The mixture was allowed to stirfor 1 h. The solids were removed by filtration and the filtrate wasconcentrated under vacuum. The crude product was purified bychromatography (SiO₂, 40:1-20:1 CH₂Cl₂:methanol) to provide4-(2-hydroxyethyl)pyridine (8.1 g, 54%) as an amber liquid.

[0801]¹H NMR (CDCl₃).

[0802] CI-MS, m/e=124 (M+1).

[0803] B. 4-(2-Hydroxyethyl)pyridine (8.1 g, 65.8 mmol) andtriethylamine (10.1 mL, 66 mmol) were dissolved in CH₂Cl₂ (100 mL) andcooled to −78° C. To this solution, tert-butyl-dimethylsilyl chloride(11.0 g, 66 mmol) was added. The mixture was allowed to warm to roomtemperature overnight. The mixture was placed directly on a bed of SiO₂and eluted with 100:0-10:1 CH₂Cl₂:methanol to provide4-[2-(tert-Butyldimethylsilyloxy)ethyl]pyridine (15.3 g, 97%).

[0804]¹H NMR (CDCl₃).

[0805] CI-MS, m/e=238 (M+1).

[0806] C. 4-[2-(tert-Butyldimethylsilyloxy)ethyl]pyridine (15.2 g, 64mmol) was dissolved in CH₂Cl₂ (200 mL). To this solution,m-chloroperbenzoic acid was added and the solution stirred at roomtemperature for 72 h. The solution was washed with aqueous NaOH (1 M)and layers separated. The water layer was extracted with CH₂Cl₂. Theorganic layers were combined, dried (K₂CO₃), filtered and concentratedunder vacuum to provide4-[2-(tert-butyldimethylsilyloxy)ethyl]pyridine-N-oxide (13.4 g, 83%).

[0807]¹H NMR (CDCl₃).

[0808] CI-MS, m/e=254 (M+1).

[0809] D. 4-[2-(tert-Butyldimethylsilyloxy)ethyl]pyridine-N-oxide (13.4g, 52.9 mmol) was dissolved in triethylamine (14.8 mL, 105 mmol).Trimethylsilyl cyanide (28.4 mL, 212 mmol) was added and the mixtureheated to 90° C. for 3 h. The mixture was allowed to cool to roomtemperature and stand overnight. The mixture was partitioned betweenwater and CH₂Cl₂. The layers were separated and water layer extractedwith CH₂Cl₂. The organic layers were combined, washed with water andbrine, dried (K₂CO₃), filtered and concentrated to provide a dark oil(14 g). The residue was purified by chromatography (SiO₂, 40:1CH₂Cl₂:ethyl acetate) to provide4-[2-(tert-butyl-dimethylsilyloxy)ethyl]-2-cyanopyridine (12.8 g, 92%)as a yellow oil.

[0810]¹H NMR (CDCl₃).

[0811] CI-MS, m/e=263 (M+1).

[0812] E. 4-[2-(tert-Butyldimethylsilyloxy)ethyl]-2-cyanopyridine (12.8g, 48.8 mmol) and a solution of tetrabutylammonium fluoride (1 M in THF,73 mL, 73 mmol) were charged to a round bottom flask and stirred at roomtemperature overnight. The mixture was diluted with water and ethylacetate. The layers were separated and the water layer extracted withethyl acetate. The organic layers were combined, washed with water andbrine, dried (Na₂SO₄), filtered and concentrated. The residue waspurified by chromatography (SiO₂, 50:1 CH₂Cl₂: methanol) to provide2-cyano-4-(2-hydroxyethyl)pyridine (4.4 g, 61%) as an off white solid.

[0813]¹H NMR (CDCl₃).

[0814] CI-MS, m/e=149 (M+1).

[0815] F. 2-Cyano-4-(2-hydroxyethyl)pyridine (4.3 g, 29 mmol) wasdissolved in pyridine (2.85 mL, 34.8 mmol) and CH₂Cl₂ (40 mL) and cooledto 0° C. To this solution, benzenesulfonyl chloride (4.5 mL, 34.8 mmol)was added and the mixture allowed to stir at room temperature overnight.The solvent was removed under vacuum and the residue was purified bychromatography (SiO₂, 20:1 CH₂Cl₂: ethyl acetate) to provide a 2:1mixture of 2-cyano-4-[2-(benzenesulfonyloxy)ethyl]pyridine and2-cyano-4-(2-chloroethyl)pyridine (6.0 g, 84%).

[0816]¹H NMR (CDCl₃).

[0817] CI-MS, m/e=167 (C₈H₇ClN₂+1) and 289 (C₁₄H₁₂N₂O₃S+1).

[0818] G. 1-Boc-piperazine (6.8 g, 36 mmol), NaI (2.7 g, 18 mmol), K₂CO₃(3.0 g, 21.6 mmol) and a mixture of2-cyano-4-[2-(benzenesulfonyloxy)ethyl]pyridine and2-cyano-4-(2-chloro-ethyl)pyridine (2:1, 4.5 g, 18 mmol) was dissolvedin DMF (50 mL) and heated to 80° C. overnight. The mixture was allowedto cool to room temperature and diluted with water and ethyl acetate.The layers were separated and the water layer extracted with ethylacetate. The organic layers were combined, washed with water and brine,dried (Na₂SO₄), filtered and concentrated under vacuum to proved a darkoil (6.0 g). The crude product was purified by chromatography (SiO₂,1000:10:1-200:10:1 CH₂Cl₂:methanol:concentrated ammonium hydroxide) toprovide 1-Boc-4-[2-(2-cyanopyridin-4-yl)ethyl]piperazine (4.0 g, 70%).

[0819]¹H NMR (CDCl₃).

[0820] TLC R_(f)=0.5 (200:10:1 CH₂Cl₂:methanol:concentrated ammoniumhydroxide).

[0821] H. 1-Boc-4-[2-(2-cyanopyridin-4-yl)ethyl]piperazine (4.0 g, 12.64mmol) was dissolved in methanol (60 mL) and cooled to 0° C. Concentratedhydrochloric acid (10.4 mL, 126 mmol) was added and the mixture stirredfor 1 h. The solvents were removed under vacuum and co-evaporated twicewith methanol, a 1:1 mixture of methanol/toluene, and finally withmethanol. The residue was dried under vacuum overnight to provide crudeproduct (4.5 g). Half of the product was dissolved in methanol,concentrated ammonium hydroxide was added, and solvents were removedunder vacuum. The residue was purified by chromatography (SiO₂, 100:10:1CH₂Cl₂:methanol:concentrated ammonium hydroxide) to provide1-[2-(2-cyanopyridin-4-yl)ethyl]piperazine (1.05 g, 69%).

[0822]¹H NMR (CDCl₃).

[0823] CI-MS, m/e=217 (M+1).

Preparation of Intermediates B-1-B-5

[0824] The following compounds were prepared according to the indicatedmethod (Method B-A, Method B-B or Method B-C) from the indicatedstarting materials, unless otherwise described.

[0825] Intermediate B-1

[0826] 1-Boc-4-[(3-pyridinyl)acetyl]piperazine.

Method B-A

[0827] 1-Boc-piperazine (12 g, 64 mmol), 3-pyridylacetic acid (8.85 g,64 mmol), and HOBt (8.64 g, 64 mmol) were dissolved in DMF. To thissolution, EDCI (14.7 g, 76.8 mmol) was added in portions. The mixturebecame homogenous and was stirred for 3 h. The mixture was diluted withwater and ethyl acetate. The layers were separated, and the aqueouslayer extracted with ethyl acetate. The organic layers were washed withwater and brine, dried (Na₂SO₄), filtered, and concentrated to provide ayellow solid. The crude product was purified by recrystallization fromhexanes:dichloromethane to provide1-Boc-4-[(3-pyridinyl)acetyl]piperazine (13.5 g, 69%) as a white solid.

[0828] 1H-NMR (CDCl₃)

[0829] CI-MS, m/e=306 (M+1)

[0830] Intermediate B-2

[0831] 1-Boc-4-[(imidazol-4-yl)acetyl]piperazine.

Method B-B

[0832] To a stirring suspension of sodium 4-imidazolylacetate (0.5 g,3.4 mmol) in DMF (25 mL) was added diethyl cyano-phosphonate (0.6 mL, 4mmol). After 5 min, Boc-piperazine (0.57 g, 3.1 mmol) was added,followed by a solution of triethylamine (0.47 mL, 3.4 mmol) in DMF (20mL). After 72 h, the solvent was removed in vacuo and the residue wasdissolved in ethyl acetate and washed with satd aq. NaHCO₃ and brine,dried with MgSO₄, filtered and concentrated to give 0.95 g of pink oil.

[0833] 1H-NMR

[0834] IS-MS, m/e 295.1 (M+1)

[0835] Intermediate B-3

[0836] 1-Boc-4-[(1-imidazolyl) acetyl]piperazine.

[0837] Preparation of Starting Materials:

[0838] 1-Boc-4-bromoacetylpiperazine.

[0839] To a stirring solution of bromoacetyl bromide (29.8 g, 148 mmol)in THF (250 mL) at 0° C. was added via an addition funnel a solution ofBoc-piperazine (25 g, 134 mmol) and triethylamine (14.9 g, 148 mmol) inTHF (75 mL). After 1 h, a few grams of ice were added and the mixturewas diluted with ethyl acetate and cold water. The layers were separatedand the organic phase was washed with 1 M aq. citric acid, brine, satdaq. NaHCO₃ and again with brine. The organic phase was then dried withMgSO₄, filtered, and concentrated in vacuo to give 38.2 g (93%) of anoff-white powder.

[0840] 1H-NMR

[0841] IS-MS, m/e 251.3 (M-C₄H₉+1)

Method B-C

[0842] To a stirring suspension of NaH (60% dispersion in mineral oil,2.34 g, 59 mmol) in THF (75 mL) was added imidazole (1.46 g, 22 mmol) insmall portions. After complete addition and complete gas evolution, asolution of 1-Boc-4-(bromoacetyl)piperazine (6 g, 19.5 mmol) in THF (40mL) was added via an addition funnel. After 2 h, the reaction wasquenched with the slow addition of water and then diluted with ethylacetate. The organic phase was washed with satd aq. NaHCO₃, followed bybrine, then dried with MgSO₄, filtered and concentrated in vacuo. Theresidue was suspended in diethyl ether with sonication, then filteredand dried to give 4.64 g (81%) of an off white powder.

[0843] 1H-NMR

[0844] IS-MS, m/e 295.2 (M+1)

[0845] Intermediate B-4

[0846] 1-Boc-4-[(1-pyrazolyl)acetyl]piperazine.

[0847] Prepared from pyrazole and 1-Boc-4-bromoacetylpiperazine usingMethod B-C.

[0848] 1H-NMR

[0849] IS-MS, m/e 295.1 (M+1)

[0850] Intermediate B-5

[0851]1-Boc-4-[(2-Benzyloxycarbonylamino)thiazol-4-ylacetyl]-piperazine.

[0852] A. (2-Aminothiazol-4-yl)acetic acid (10 g, 63.2 mmol) wasdissolved in 1,4-dioxane (100 mL) and aqueous NaOH (6 M, 100 mL), andthe solution was cooled to 0° C. Benzyl chloro-formate (20 mL, 82.2mmol) was added, and the mixture was stirred at room temperatureovernight. The mixture was diluted with diethyl ether, and the layersseparated. The water layer was cooled to 0° C., and the pH adjusted toapproximately 4 with aqueous HCl (6 M). The white precipitate formed wascollected by vacuum filtration, washed with water and diethyl ether, anddried under vacuum to provide(2-benzyloxycarbonylaminothiazol-4-yl)acetic acid (7.5 g, 41%).

[0853]¹H NMR (DMSO-d₆).

[0854] CI-MS, m/e=293 (M+1).

[0855] B. Using methods substantially equivalent to those described inMethod B-A, the title compound was prepared from(2-benzyloxycarbonylaminothiazol-4-yl) acetic acid and 1-Boc-piperazine(95%).

[0856]¹H NMR (CDCl₃).

[0857] CI-MS, m/e=461 (M+1).

Preparation of Intermediates C-1-C-28

[0858] The following compounds were prepared according to the indicatedmethod (Method C-A, Method C-B, Method C-C or Method C-D) from theindicated starting materials, unless otherwise described.

[0859] Intermediate C-1

[0860] 1-(Boc-D-Phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine.

Method C-A

[0861] D-Boc-Phenylglycine (8.4 g, 33.3 mmol) and1-[2-(4-pyridinyl)ethyl]piperazine hydrochloride (10 g, 33.3 mmol) weredissolved in DMF (500 mL) and cooled to approximately −15° C. in anice-methanol bath. Diethyl cyanophosphonate (5.5 mL, 36.6 mmol) wasslowly added to the mixture. Triethylamine (18.6 mL, 133.2 mmol) wasadded dropwise to the solution. The mixture was stirred at −15° C. for 2h and was allowed to gradually warm to-room temperature overnight. Themixture was diluted with ethyl acetate and water. The layers wereseparated, and the water layer extracted with ethyl acetate. The organiclayers were combined, washed with brine, dried (Na₂SO₄), filtered, andconcentrated under vacuum. The crude product was filtered through a plugof silica gel (1.2 kg) using 1:1 hexanes:ethyl acetate as eluent toprovide 1-(Boc-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine(10.6 g, 75%) as a light yellow oil.

[0862] 1H-NMR (CDCl₃)

[0863] CI-MS, m/e=425 (M+1)

[0864] Intermediate C-2

[0865] 1-(Boc-D-Phenylglycinyl)-4-[2-(2-pyridinyl)ethyl]piperazine.

[0866] Prepared from Boc-D-phenylglycine and1-[2-(2-pyridinyl)-ethyl]piperazine using Method C-A.

[0867] 1H-NMR(CDCl₃)

[0868] CI-MS, m/e=425 (M+1)

[0869] Intermediate C-3

[0870] 1-(Boc-D-Phenylglycinyl)-4-[2-(2-pyrazinyl)ethyl]piperazine.

[0871] Prepared from Boc-D-phenylglycine and1-[2-(2-pyrazinyl)-ethyl]piperazine using Method C-A.

[0872] 1H-NMR(CDCl₃)

[0873] CI-MS, m/e=426 (M+1)

[0874] Intermediate C-4

[0875] 1-(Boc-D-Phenylglycinyl)-4-[2-(3-pyridazinyl)ethyl]piperazine.

[0876] Prepared from Boc-D-phenylglycine and1-[2-(3-pyridazinyl)ethyl]piperazine using Method C-A.

[0877] 1H NMR (CDCl₃)

[0878] TLC R_(f)=0.65 (100:10:1 CH₂Cl₂:MeOH:NH₄OH, SiO₂, Analtech No.02521)

[0879] Intermediate C-5

[0880] 1-(Boc-D-Phenylglycinyl)-4-[2-(3-pyridinyl)ethyl]piperazine.

[0881] Prepared from Boc-D-phenylglycine and1-[2-(3-pyridinyl)-ethyl]piperazine using Method C-A.

[0882] 1H-NMR(CDCl₃)

[0883] CI-MS, m/e=425 (M+1)

[0884] Intermediate C-6

[0885] 1-(Boc-D-Phenylglycinyl)-4-[2-(4-imidazolyl)ethyl]piperazine

[0886] Prepared from Boc-D-phenylglycine and1-[2-(4-imidazol-yl)ethyl]piperazine using Method C-A.

[0887] 1H-NMR

[0888] IS-MS, m/e 414.2 (M+1)

[0889] Intermediate C-7

[0890] 1-(Boc-D-Phenylglycinyl)-4-[2-(4-pyrazolyl)ethyl]piperazine.

[0891] Prepared from Boc-D-phenylglycine and1-[2-(4-pyrazolyl)-ethyl]piperazine using Method C-A.

[0892] 1H-NMR

[0893] IS-MS, m/e 414.2 (M+1)

[0894] Intermediate C-8

[0895] 1-(Boc-D-Phenylglycinyl)-4-[2-(1-imidazolyl)ethyl]piperazine.

[0896] Prepared from Boc-D-phenylglycine and1-[2-(1-imidazolyl)ethyl]piperazine using Method C-A.

[0897] 1H-NMR

[0898] IS-MS, m/e 414.2 (M+1)

[0899] Intermediate C-9

[0900] 1-(Boc-D-Phenylglycinyl)-4-[2-(1-pyrazolyl)ethyl]piperazine.

[0901] Prepared from Boc-D-phenylglycine and1-[2-(1-pyrazolyl)-ethyl]piperazine using Method C-A.

[0902] 1H-NMR

[0903] IS-MS, m/e 414.2 (M+1)

[0904] Intermediate C-10

[0905]1-[Boc-D,L-(Pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]-piperazine.

Method C-B

[0906] To a stirring solution of ethyl Boc-D,L-(pyridin-2-yl)-glycine(16.3 g, 58.2 mmol) in 1,4-dioxane (100 ml) was added a solution of LiOHhydrate (2.68 g, 64 mmol) in water (100 mL). After 2 h, another solutionof LiOH hydrate (1.34 g, 32 mmol) in water (50 mL) was added. Afteranother 2 h, the solvent was evaporated in vacuo to give 13.56 g ofoff-white solid.

[0907] A portion of the solid (3 g, 11.6 mmol) was dissolved in DMF (75mL) and cooled to 0° C. To this solution was added diethylcyanophosphonate (2.3 g, 13.9 mmol), N,N-diisopropylethylamine (6 g,46.4 mmol) and then 1-[2-(4-pyridyl)ethyl]piperazine hydrochloride (3.8g, 12.8 mmol), and the reaction was allowed to slowly warm to roomtemperature overnight. The next morning, the solvents were removed invacuo and the residue was dissolved in ethyl acetate and washed withsatd aq. NaHCO₃ and brine, then dried with Na₂SO₄, filtered, andconcentrated in vacuo. The residue was then dissolved in a minimalvolume of dichloromethane and chromatographed over silica gel, elutingwith a step gradient of 2% through 10% methanol (with 2 N NH₃) indichloromethane. The product containing fractions were combined andconcentrated in vacuo to give 2.31 g (47%) of an off-white foam.

[0908] 1H-NMR

[0909] IS-MS, m/e 426.3 (M+1)

[0910] Intermediate C-11

[0911]1-[Boc-D,L-(2-Methoxyphenyl)glycinyl]-4-[2-(4-pyridinyl)-ethyl]piperazine.

Method C-C

[0912] To a stirring solution of Boc-D,L-(2-methoxyphenyl)-glycine (2 g,7.1 mmol) and 1-[2-(4-pyridinyl)ethyl]piperazine trihydrochloride (2.4g, 7.8 mmol) in DMF (50 mL), was added HOBt (1.06 g, 7.8 mmol), andtriethylamine (4.96 mL, 35.6 mmol) followed by DCC (1.61 g, 7.8 mmol).After stirring overnight at room temperature, the mixture was filtered;and the filtrate was concentrated in vacuo. The residue was dissolved inethyl acetate and washed with satd aq. NaHCO₃ followed by brine, thendried with MgSO₄, filtered and concentrated in vacuo. The residue wasthen dissolved in a minimum amount of dichloromethane andchromatographed over silica gel, eluting with a step gradient ofdichloromethane through 10% (2 N NH₃/methanol) in dichloromethane. Theproduct containing fractions were combined and concentrated in vacuo togive 2.5 g (77%) of the title compound.

[0913] 1H-NMR

[0914] IS-MS, m/e 455.1 (M+1)

[0915] Intermediate C-12

[0916] 1-(Boc-D-Phenylglycinyl)-4-[2-(thiazol-2-yl)ethyl]piperazine.

[0917] Prepared from 1-[2-(thiazol-2-yl)ethyl)piperazine dihydrochlorideand Boc-D-phenylglycine using Method C-A, except using dichloromethanein place of DMF (80%).

[0918]¹H NMR (CDCl₃).

[0919] CI-MS, m/e=431 (C₂₂H₃₀N₄O₃S+1).

[0920] Intermediate C-13

[0921]1-(Boc-D-Phenylglycinyl)-4-[2-(2-benzyloxycarbonylamino-thiazol-4-yl)ethyl]piperazine.

[0922] Prepared from1-[2-(2-benzyloxycarbonylaminothiazol-4-yl)ethyl]piperazinehydrochloride and Boc-D-phenylglycine using Method C-A (76%).

[0923]¹H NMR (CDCl₃).

[0924] APCI-MS, m/e=580 (M+1).

[0925] Intermediate C-14

[0926]1-(Boc-D-Phenylglycinyl)-4-[2-(3-fluoropyridin-4-yl)ethyl]-piperazine.

[0927] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(3-fluoro-pyridin-4-yl)ethyl]piperazineusing Method C-A, except using N,N-diisopropylethylamine in place oftriethylamine and dichloromethane in place of DMF (89%).

[0928]¹H NMR (CDCl₃).

[0929] APCI-MS, m/e=443 (M+1).

[0930] Intermediate C-15

[0931]1-(Boc-D-Phenylglycinyl)-4-[2-(2-cyanopyridin-4-yl)ethyl]-piperazine.

Method C-D

[0932] 1-[2-(2-cyanopyridin-4-yl)ethyl]piperidine (1.0 g, 4.6 mmol) andBoc-D-phenylglycine (1.39 g, 4.63 mmol) were dissolved in CH₂Cl₂ (20 mL)and cooled to −10° C. To this solution, diethyl cyanophosphonate (0.94mL, 4.63 mmol) was added, followed by a solution of triethylamine (0.97mL, 6.9 mmol) in CH₂Cl₂ (10 mL). The mixture was allowed to slowly warmto room temperature overnight. The mixture was diluted with water andthe layers separated. The water layer was extracted with CH₂Cl₂. Theorganic layers were combined, washed with brine, dried (Na₂SO₄),filtered and concentrated under vacuum to provide crude product (3.0 g).The crude product was purified by chromatography (SiO₂,1000:10:1-400:10:1 CH₂Cl₂:methanol:concentrated ammonium hydroxide) toprovide1-(Boc-D-phenylglycinyl)-4-[2-(2-cyanopyridin-4-yl)-ethyl]piperazine(1.61 g, 78%).

[0933]¹H NMR (CDCl₃).

[0934] CI-MS, m/e=450 (M+1).

[0935] Intermediate C-16

[0936]1-[Boc-D,L-(2-Chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[0937] Prepared from Boc-D,L-(2-chlorophenyl)glycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, with EDCI inplace of DCC and HOAt in place of HOBt.

[0938] 1H NMR

[0939] IS-MS, m/e 451.0 (M+1)

[0940] Intermediate C-17

[0941]1-[Boc-D,L-(Quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[0942] Prepared from Boc-D,L-(quinolin-8-yl)glycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, with EDCI inplace of DCC and HOAt in place of HOBt.

[0943] 1H NMR

[0944] Intermediate C-18

[0945]1-[Boc-D,L-(2-Trifluoromethylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[0946] Prepared from Boc-D,L-(2-trifluoromethylphenyl)glycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, with DIEA inplace of TEA.

[0947] 1H NMR

[0948] IS-MS, m/e 485.0 (M+1)

[0949] Intermediate C-19

[0950]1-[Boc-D-Cyclopentylglycinyl]-4-(1-methylpiperidin-4-yl)-piperazine.

[0951] Prepared from Boc-D-cyclopentylglycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, with EDCI inplace of DCC and HOAt in place of HOBt.

[0952] 1H NMR

[0953] IS-MS, m/e 409.3 (M+1)

[0954] Intermediate C-20

[0955]1-[Boc-D-Cyclohexylglycinyl]-4-(1-methylpiperidin-4-yl)-piperazine.

[0956] Prepared from Boc-D-cyclohexylglycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, with EDCI inplace of DCC and HOAt in place of HOBt.

[0957] 1H NMR

[0958] IS-MS, m/e 423.3 (M+1)

[0959] Intermediate C-21

[0960] 1-[Cbz-D-Phenylglycinyl]-4-(2-phenethyl)piperazine.

[0961] Prepared from 4-(Cbz-D-phenylglycinyl)piperazine andphenylacetaldehyde using Method I-A, with sodium triacetoxyborohydridein place of sodium cyanoborohydride and dichloroethane in place ofmethanol (71%).

[0962]¹H NMR (CDCl₃).

[0963] APCI-MS, m/e=458 (M+1).

[0964] Intermediate C-22

[0965]1-[Boc-D,L-(thiazol-2-yl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazine.

[0966] Prepared from ethyl Boc-D,L-thiazol-2-ylglycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-B.

[0967] IS-MS, m/e 424.0 (M+1)

[0968] Intermediate C-23

[0969]1-[Boc-D,L-(Benzo[b]thiophen-3-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[0970] Prepared from Boc-D,L-(benzo[b]thiophen-3-yl)glycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, substituting EDCIfor DCC, N,N-diisopropylamine for triethylamine, and substitutingdichloromethane for DMF.

[0971] 1H-NMR

[0972] LCMS m/z 473.4 (M+1)

[0973] Intermediate C-24

[0974]1-[Boc-D,L-(Naphthalen-1-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[0975] Prepared from Boc-D,L-naphthalen-1-ylglycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, substituting EDCIfor DCC and substituting N,N-diisopropylamine for triethylamine.

[0976] 1H-NMR

[0977] IS-MS, m/e 467.1 (M+1)

[0978] Intermediate C-25

[0979]1-[Boc-D,L-(2-Methylsulfonylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[0980] Prepared from Boc-D,L-(2-methylsulfonylphenyl)glycine and1-(1-methylpiperidin-4-yl)piperazine using Method C-C, substitutingEDCI/HOAt for DCC/HOBt and substituting N,N-diisopropylethylamine fortriethylamine.

[0981] 1H-NMR

[0982] IS-MS, m/e 495 (M+1)

[0983] Analysis for C₂₄H₃₈N₄O₅S: Calcd: C, 58.27; H, 7.74; N, 11.32;Found: C, 58.05; H, 7.63; N, 11.43.

[0984] Intermediate C-26

[0985]1-[Boc-D,L-Thiazol-5-ylglycinyl]-4-[2-(pyridin-4-yl)ethyl]-piperazine.

[0986] To a solution of Boc-D,L-thiazol-5-ylglycine (1.33 g, 5.15 mmol),HOAt (772 mg, 5.67 mmol), 1-[2-(pyridin-4-yl)-ethyl]piperazinedihydrochloride (1.55 g, 5.15 mmol) and triethylamine (1.58 mL, 11.3mmol) in DMF (41 mL) was added EDCI (1.09 g, 5.67 mmol), and the mixturestirred at room temperature for 18 h. The solvent was removed in vacuo,the residues taken up in chloroform: isopropyl alcohol (2:1) and washedwith water, satd aqueous sodium bicarbonate, dried (MgSO₄) andconcentrated in vacuo to an orange-brown oil. The crude reaction productthus obtained was carried on to the next step without furtherpurification.

[0987] Intermediate C-27

[0988]1-[Boc-D,L-(2-Methylthiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazine.

[0989] Prepared from Boc-D,L-(2-methylthiazol-4-yl)glycine and1-[2-(pyridin-4-yl)ethyl]piperazine dihydrochloride using proceduressubstantially equivalent to those described for the preparation ofIntermediate C-26.

[0990] Intermediate C-28

[0991]1-[Boc-D,L-(2-Benzyloxycarbonylaminothiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazine.

[0992] Prepared fromBoc-D,L-(2-benzyloxycarbonylaminothiazol-4-yl)glycine and1-[2-(pyridin-4-yl)ethyl]piperazine dihydrochloride using proceduressubstantially equivalent to those described for the preparation ofIntermediate C-26.

Preparation of Intermediates D-1-D-28

[0993] The following compounds were prepared according to the indicatedmethod (Method D-A, Method D-B, Method D-C, Method D-D or Method D-F)from the indicated starting material, unless otherwise described.

[0994] Intermediate D-1

[0995] 1-(D-Phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazineHydrochloride.

Method D-A

[0996] 1-(Boc-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]-piperazine (13g, 30.6 mmol) and anisole (50 mL) were dissolved in methanol and cooledto 0° C. Concentrated hydrochloric acid (40 mL, 300 mmol) was addeddropwise to the solution, and the mixture allowed to warm to roomtemperature. The mixture stirred for 1 h, and the solvent and anisolewere removed under vacuum. The residue was suspended in diethyl etherand sonicated for 1 h. The solid product was filtered and dried undervacuum (0.5 torr, 66 Pa at 50-60° C.) to give1-(D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine hydrochloride(11.8 g, 89%) as a white, hygroscopic solid.

[0997] 1H NMR (CD₃OD)

[0998] API-MS, m/e=325 (M+1)

[0999] Intermediate D-2

[1000] 1-(D-Phenylglycinyl)-4-[2-(2-pyridinyl)ethyl]piperazine.

[1001] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(2-pyridin-yl)ethyl]piperazine usingMethod D-A.

[1002] 1H NMR (CD₃OD)

[1003] API-MS, m/e=325 (M+1)

[1004] Intermediate D-3

[1005] 1-(D-Phenylglycinyl)-4-[2-(2-pyrazinyl)ethyl]piperazine.

[1006] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(2-pyrazin-yl)ethyl]piperazine usingMethod D-A.

[1007] 1H NMR (CD₃OD)

[1008] API-MS, m/e=326 (M+1)

[1009] Intermediate D-4

[1010] 1-(D-Phenylglycinyl)-4-[2-(3-pyridazinyl)ethyl]piperazine.

[1011] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(3-pyridazinyl)ethyl]piperazine usingMethod D-A.

[1012] 1H-NMR (CD₃OD)

[1013] IS-MS, m/e 326 (M+1)

[1014] Intermediate D-5

[1015] 1-(D-Phenylglycinyl)-4-[2-(3-pyridinyl)ethyl]piperazine.

[1016] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(3-pyridin-yl)ethyl]piperazine usingMethod D-A.

[1017] 1H NMR (CD₃OD)

[1018] API-MS, m/e=325 (M+1)

[1019] Intermediate D-6

[1020] 1-(D-Phenylglycinyl)-4-[2-(4-imidazolyl)ethyl]piperazine.

[1021] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(4-imidazolyl)ethyl]piperazine usingMethod D-A.

[1022] 1H-NMR

[1023] IS-MS, m/e 314.1 (M+1)

[1024] Intermediate D-7

[1025] 1-(D-Phenylglycinyl)-4-[2-(4-pyrazolyl)ethyl]piperazine.

[1026] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(4-pyrazol-yl)ethyl]piperazine usingMethod D-A.

[1027] 1H-NMR

[1028] IS-MS, m/e 314.3 (M+1)

[1029] Intermediate D-8

[1030] 1-(D-Phenylglycinyl)-4-[2-(1-imidazolyl)ethyl]piperazine.

[1031] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(1-imidazolyl)ethyl]piperazine usingMethod D-A.

[1032] 1H-NMR

[1033] IS-MS, m/e 314.1 (M+1)

[1034] Intermediate D-9 (PD7-H7C-045, -046)

[1035] 1-(D-Phenylglycinyl)-4-[2-(1-pyrazolyl)ethyl]piperazine.

[1036] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(1-pyrazol-yl)ethyl]piperazine usingMethod D-A.

[1037] 1H-NMR

[1038] IS-MS, m/e 314.1 (M+1)

[1039] Intermediate D-10

[1040]1-[D,L-(Pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]-piperazine.

Method D-B

[1041] To a stirring solution of1-[Boc-D,L-(pyridin-2-yl)-glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazine(2.31 g, 5.4 mmol) in dichloromethane (45 mL) was added TFA (5 mL).After 6 h, the solvents were removed in vacuo. The residue waspartitioned between ethyl acetate and satd aq. NaHCO₃, and the layerswere separated. The aqueous phase was extracted with 50% ethylacetate/dichloromethane, then 5% methanol/dichloromethane. The combinedorganic extracts were dried with MgSO₄, filtered and concentrated togive 1.66 g (94%) of the title compound.

[1042] 1H-NMR

[1043] IS-MS, m/e 326.1 (M+1)

[1044] Intermediate D-11

[1045]1-[D,L-(2-Methoxyphenyl)glycinyl]-4-[2-(1-pyrazolyl)ethyl]-piperazine.

[1046] Prepared from1-[Boc-D,L-(2-methoxyphenyl)glycinyl]-4-(2-(1-pyrazolyl)ethyl]piperazineusing Method D-B.

[1047] 1H-NMR

[1048] IS-MS, m/e 355.1 (M+1)

[1049] Intermediate D-12

[1050] 1-(D-Phenylglycinyl)-4-[2-(thiazol-2-yl)ethyl]piperazineTrihydrochloride.

[1051] Prepared from1-[((Boc-D-phenylglycinyl)-4-[2-(thiazol-2-yl)ethyl]piperazine usingmethods substantially equivalent to those described in Method D-A (80%).

[1052]¹H NMR (CD₃OD).

[1053] CI-MS, m/e=331 (C₁₇H₂₂N₄OS+1).

[1054] Intermediate D-13

[1055]1-(D-Phenylglycinyl)-4-[2-(2-benzyloxycarbonylaminothiazol-4-yl)ethyl]piperazineTrihydrochloride.

Method D-C

[1056]1-(Boc-D-Phenylglycinyl)-4-[2-(2-benzyloxycarbonylamino-thiazol-4-yl)ethyl]piperazine(520 mg, 0.898 mmol) was dissolved in ethyl acetate (10 mL) and anisole(1 mL). The mixture was cooled to 0° C., and a saturated solution of HClin ethyl acetate was added. The mixture was allowed to warm to roomtemperature and stirred for 4 h. The solvent was removed under vacuum toprovide the title compound as a white solid (530 mg, quantitative).

[1057]¹H NMR (CD₃OD).

[1058] APCI-MS, m/e=480 (C₂₅H₂₉N₅O₃S+1).

[1059] Intermediate D-14

[1060] 1-(D-Phenylglycinyl)-4-[2-(3-fluoropyridin-4-yl)ethyl]-piperazineTrihydrochloride.

[1061] Prepared from1-(Boc-D-phenylglycinyl)-4-[2-(3-fluoro-pyridin-4-yl)ethyl]piperazineusing a method substantially equivalent to Method D-A (98%).

[1062]¹H NMR (CD₃OD).

[1063] APCI-MS, m/e=353 (C₁₉H₂₃FN₄O+1).

[1064] Intermediate D-15

[1065] 1-(D-Phenylglycinyl)-4-[2-(2-cyanopyridin-4-yl)ethyl]-piperazineTrihydrochloride.

Method D-D

[1066]1-(Boc-D-phenylglycinyl)-4-[2-(2-cyanopyridin-4-yl)-ethyl]piperazine(580 mg, 1.29 mmol) and anisole (5.0 g, 4.64 mmol) were dissolved inmethanol (10 mL) and cooled to −15° C. To this solution, concentrated (6N) hydrochloric acid (1.2 mL, 11.6 mmol) was added, and the mixturestirred at −10° C. for 1 hour. The solvents were removed under vacuumand the residue co-evaporated with methanol, methanol and toluene, andmethanol and ethyl acetate to provide1-(D-phenylglycinyl)-4-(2-(2-cyanopyridin-4-yl)ethyl]piperazinetrihydrochloride (600 mg, 100%) as an off white solid.

[1067]¹H NMR (CD₃OD).

[1068] CI-MS, m/e=350 (M+1).

[1069] Intermediate D-16

[1070]1-[D,L-(2-Chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazine.

[1071] Prepared from1-[Boc-D,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineusing Method D-E.

[1072] 1H NMR

[1073] IS-MS, m/e (M+1)

[1074] Intermediate D-17

[1075]1-[D,L-(Quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazine.

Method D-E

[1076] To a stirring solution of1-[Boc-D,L-(quinolin-8-yl)-glycinyl]-4-(1-methylpiperidin-4-yl)piperazine(0.53 g, 1.13 mmol) and anisole (0.62 mL, 5.67 mmol) in dichloromethane(22 mL) was added TFA (2.2 mL). After 4 h, the solvents were removed invacuo, and the residue was dissolved in methanol and loaded onto an SCXcolumn (pretreated with 5% acetic acid in methanol and washed withmethanol). The column was washed with methanol and then the product waseluted with 30% (2 N ammonia/methanol) in dichloromethane. The productcontaining fractions were combined and concentrated in vacuo to giveapproximately 0.4 g (quantitative) of the title compound.

[1077] 1H NMR

[1078] Intermediate D-18

[1079]1-[D,L-(2-Trifluoromethylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1080] Prepared from1-[Boc-D,L-(2-trifluoromethylphenyl)-glycinyl]-4-(1-methylpiperidin-4-yl)piperazineusing Method D-E.

[1081] 1H NMR

[1082] IS-MS, m/e 385.1 (M+1)

[1083] Intermediate D-19

[1084] 1-[D-Cyclopentylglycinyl]-4-(1-methylpiperidin-4-yl)-piperazine.

[1085] Prepared from1-[Boc-D-cyclopentylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine usingMethod D-E.

[1086] 1H NMR

[1087] IS-MS, m/e 309.2 (M+1)

[1088] Intermediate D-20

[1089] 1-[D-Cyclohexylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1090] Prepared from1-[Boc-D-cyclohexylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine usingMethod D-E.

[1091] 1H NMR

[1092] IS-MS, m/e 323.3 (M+1)

[1093] Intermediate D-21

[1094] 1-[D-Phenylglycinyl]-4-(2-phenethyl)piperazine.

Method D-F

[1095] 1-(Cbz-D-Phenylglycinyl)-4-(2-phenethyl)piperazine (1.66 g; 3.63mmol) was dissolved in methanol (46 mL) with 10% Pd/C (394 mg) andreaction mixture was subjected to a balloon of hydrogen for 15 h. Only50% conversion was observed; so the catalyst was filtered throughdiatomaceous earth, and the mixture was re-subjected to the sameconditions for 17 h. The catalyst was filtered through diatomaceousearth, and the solvent was removed under vacuum to give the titlecompound (1.03 g; 88%).

[1096]¹H NMR (CDCl₃).

[1097] APCI-MS, m/e=324 (M+1).

[1098] Intermediate D-22

[1099]1-[D,L-(Thiazol-2-yl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazineTrihydrochloride.

[1100] Prepared from1-[Boc-D,L-(thiazol-2-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineusing Method A-A. In this procedure, saturated HCl in 1,4-dioxane wasused in place of saturated HCl in methanol. Concentration of thereaction mixture provided the title compound as a crude residue that wasused directly without purification.

[1101] Intermediate D-23

[1102]1-[D,L-(Benzo[b]thiophen-3-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineTrihydrochloride.

[1103] Prepared from1-[Boc-D,L-(benzo[b]thiophen-2-yl)-glycinyl]-4-(1-methylpiperidin-4-yl)piperazineusing Method A-A, Part B. In this procedure, saturated HCl in1,4-dioxane was used in place of saturated HCl in methanol.

[1104] LCMS m/z 373.5 (M+1)

[1105] Intermediate D-24

[1106]1-[D,L-(Naphthalen-1-yl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazineTrihydrochloride.

[1107] Prepared from1-[Boc-D,L-(naphthalen-1-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineusing Method A-A, Part B. In this procedure, saturated HCl in1,4-dioxane was used in place of saturated HCl in methanol.

[1108] 1H-NMR

[1109] IS-MS m/e 367.0 (M+1)

[1110] Intermediate D-25

[1111]1-D,L-(2-Methylsulfonylphenyl)glycinyl-4-(1-methylpiperidin-4-yl)piperazine.

[1112] Prepared from1-[Boc-D,L-(2-methylsulfonylphenyl)-glycinyl]-4-(1-methylpiperidin-4-yl)piperazineusing Method D-D.

[1113] 1H-NMR

[1114] IS-MS, m/e 395 (M+1)

[1115] Intermediate D-26

[1116]1-[D,L-(Thiazol-5-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]-piperazine.

[1117] To a stirred solution of crude1-[D,L-(thiazol-5-yl)-glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazine(circa 5.15 mmol) and anisole (11.2 mL) in dichloromethane (42 mL) atroom temperature was added TFA (10.5 mL), and the mixture stirred atroom temperature for 16 h before concentrating in vacuo. The product wasisolated using SCX ion exchange chromatography.

[1118] NMR

[1119] Intermediate D-27

[1120]1-[D,L-(2-Methylthiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)-ethyl]piperazine.

[1121] Prepared from1-[Boc-D,L-(2-methylthiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazineusing procedures substantially equivalent to those described in thepreparation of Intermediate D-26.

[1122] NMR

[1123] Intermediate D-28

[1124]1-[D,L-(2-Aminothiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)-ethyl]piperazine.

[1125] A stirred solution of1-(Boc-D,L-2-benzyloxycarbonyl-aminothiazol-4-ylglycinyl)-4-[2-(pyridin-4-yl)ethyl]piperazine(crude, circa 4.2 mmol) in a mixture of HBr-acetic acid (50%, 35 mL) andacetic acid (70 mL) was heated at 60° C. for 6 h, cooled and thenconcentrated in vacuo. The title product was isolated using SCX ionexchange chromatography.

[1126] NMR

Preparation of Intermediates E

[1127] The following compounds were prepared according to the indicatedmethod (Method E-A) from the indicated starting material, unlessotherwise described.

[1128] Intermediate E-1

[1129] 1-Boc-4-(Cbz-D-phenylglycinyl)piperazine.

Method E-A

[1130] D-Cbz-phenylglycine (58.0 g, 203 mmol) and 1-Boc-piperazine (41.7g, 224 mmol) were dissolved in DMF (1 L) and cooled to approximately−15° C. in an ice-methanol bath. Diethyl cyanophosphonate (37.0 mL, 244mmol) was slowly added to the mixture. Triethylamine (59.4 mL, 426 mmol)was added dropwise to the solution. The mixture was stirred at −15° C.for 2 h and was allowed to gradually warm to room temperature overnight.The mixture was diluted with ethyl acetate and water. The layers wereseparated, and the water layer extracted with ethyl acetate. The organiclayers were combined, washed with 10% citric acid (2×500 mL) and brine,dried (Na₂SO₄), filtered and concentrated under vacuum. The crudeproduct was filtered through a plug of silica gel (1.2 kg) using 1:1hexanes:ethyl acetate as eluent to provide1-Boc-4-(Cbz-D-phenylglycinyl)piperazine (69.9 g, 76%) as a colorlessoil.

[1131] 1H-NMR(CDCl₃)

[1132] API-MS, m/e=454 (M+1)

Preparation of Intermediates F

[1133] The following compounds were prepared according to the indicatedmethod (Method F-A) from the indicated starting material, unlessotherwise described.

[1134] Intermediate F-1

[1135] 1-Boc-4-(D-phenylglycinyl)piperazine.

Method F-A

[1136] 1-Boc-4-(Cbz-D-phenylglycinyl)piperazine (69.5 g, 153 mmol) wasdissolved in ethanol (500 mL). The mixture was degassed with nitrogenand 10% Pd/C (6.8 g) was added. Hydrogen was bubbled through the mixturefor 1 h, and it was maintained under a hydrogen atmosphere for 16 h. ThePd/C was removed by filtration through cellulose. The filter cake wasrinsed with ethanol and ethyl acetate. The filtrate was concentratedunder vacuum to give 1-Boc-4-(D-phenylglycinyl)piperazine (45.3 g, 93%)as a light yellow solid.

[1137] 1H-NMR(CDCl₃)

[1138] API-MS, m/e=320 (M+1)

Preparation of Intermediates G

[1139] The following compounds were prepared according to the indicatedmethod (Method G-A) from the indicated starting material, unlessotherwise described.

[1140] Intermediate G-1

[1141] 1-Boc-4-(4-Methoxybenzoyl-D-phenylglycinyl)piperazine.

Method G-A

[1142] 1-Boc-4-(D-phenylglycinyl)piperazine (42.0 g, 131.5 mmol) wasdissolved in 1,4-dioxane (420 mL) and water (210 mL) and was cooled to10° C. Potassium carbonate (36.4 g, 263 mmol) was added, followed byp-methoxybenzoyl chloride (24.7 g, 144 mmol). The mixture stirred atroom temperature overnight. The mixture was diluted with water and ethylacetate. The layers were separated and the water layer extracted withethyl acetate. The organic layers were combined, washed with brine,dried, filtered and concentrated to provide1-Boc-4-(4-methoxybenzoyl-D-phenylglycinyl)piperazine (58.7 g, 98%) asan off-white solid.

[1143] 1H-NMR(CDCl₃)

[1144] API-MS, m/e=454 (M+1)

Preparation of Intermediates H

[1145] The following compounds were prepared according to the indicatedmethod (Method H-A) from the indicated starting material, unlessotherwise described.

[1146] Intermediate H-1

[1147] 1-(4-Methoxybenzoyl-D-phenylglycinyl)piperazine Trifluoroacetate.

Method H-A

[1148] 1-Boc-4-(4-Methoxybenzoyl-D-phenylglycinyl)piperazine (20.0 g,44.1 mmol) was dissolved in dichloromethane (50 mL) and anisole (20 mL).To this vigorously stirred mixture was added trifluoroacetic acid (50mL). The mixture was stirred for 25 min at room temperature. Thesolvents were removed under vacuum. The residue was triturated in etherand sonicated for 60 min. The solid was collected by filtration anddried in a vacuum pistol overnight to provide1-(4-methoxybenzoyl-D-phenylglycinyl)piperazine trifluoroacetate (18.2g, 88%) as a light yellow solid.

[1149] 1H-NMR(CD₃OD)

[1150] API-MS, m/e=354 (M+1)

PREPARATION OF EXAMPLES

[1151] The following examples of formula (I) were prepared according tothe indicated method (Method I-A, Method I-B, Method I-C, Method I-D,Method I-E, Method I-F or Method I-G) from the indicated startingmaterials, unless otherwise described.

Example 1

[1152] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-phenethylpiperazine.

Method I-A

[1153] To a stirring solution of1-(4-methoxybenzoyl-D-phenylglycinyl)piperazine (0.05 g, 0.14 mmol) inmethanol (1 mL) was added phenylacetaldehyde (0.17 mL, 1.4 mmol),followed by acetic acid (0.05 mL, 0.87 mmol) and then sodiumcyanoborohydride (0.014 g, 0.21 mmol). After 2 h, the solution wasloaded onto an SCX column, which was pretreated with 5% aceticacid/methanol. The column was washed with methanol and then the productwas eluted with 10/1 dichloromethane:(2 N NH₃ in methanol). The productcontaining fractions were combined and concentrated to give 63 mg ofthick oil (90% pure by analytical HPLC). The crude product was dissolvedin a minimal volume of dichloromethane and chromatographed over silicagel, eluting with dichloromethane, followed by ethyl acetate, followedby a gradient of 2% through 10% (2 N NH₃/methanol) in dichloromethane.The product containing fractions were combined and concentrated to give0.022 g (34%) of the title compound.

[1154] 1H-NMR

[1155] IS-MS, m/e 458.0 (M+1)

[1156] HPLC Analysis (Method A): 100% t_(r)=27.44 min.

Example 2

[1157]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)-ethyl]piperazineDiydrochloride.

Method I-B

[1158] 1-(D-Phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazinetrihydrochloride (1.0 g, 2.31 mmol) and potassium carbonate (2.0 g,144.4 mmol) were dissolved in 1,4-dioxane (5 mL) and water (1 mL). Tothis solution, p-anisoyl chloride (650 μL, 4.62 mmol) was added. Themixture stirred at room temperature for 3 h. The mixture was dilutedwith water, and the mixture extracted with ethyl acetate. The organiclayers were combined, washed with brine, dried (Na₂SO₄), filtered andconcentrated. The residue was dissolved in methanol and loaded onto anSCX column (10 g, pretreated with 5% acetic acid in methanol and washedwith methanol). The by-products were eluted with methanol (about 20 mL),and desired product eluted with saturated ammonia in methanol. Theproduct was further purified by column chromatography (SiO₂, CH₂Cl₂:CMA20:1 to 9:1 gradient). The product was dissolved in methanol, and HCl indiethyl ether was added to provide1-(4-methoxybenzoyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazinehydrochloride (370 mg, 37%) as an off white solid.

[1159] 1H-NMR(CDCl₃)

[1160] CI-MS, m/e=459 (M+1)

[1161] Analysis for C₂₇H₃₀N₄O₃.2.2 HCl.1.1 H₂O.0.4 NH₄Cl: Calcd: C,55.91; H, 6.26; N, 10.63; Found: C, 56.04; H, 6.55; N, 10.46.

[1162] HPLC Analysis (Method B): 99.7%, t_(R)=10.98 min.

Example 3

[1163]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)-ethyl]piperazine.

Method I-C

[1164] 1-(D-Phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine (1.0 g,2.31 mmol), 6-carboxyindole (371 mg, 2.31 mmol), HOBt (312 mg, 2.31mmol), Et₃N (1.3 mL, 9.24 mmol), and DCC (620 mg, 3.00 mmol) werestirred in DMF at room temperature overnight. The precipitate wasremoved by filtration, and the filtrate concentrated under vacuum to athick paste. The residue was dissolved in methanol and purified by ionexchange chromatography (SCX resin, methanol then saturated NH₃ inmethanol) to provide the crude product as a brown solid. The crudeproduct was purified by chromatography (SiO₂, 20:1 CH₂Cl₂:CMA to 6:1CH₂Cl₂:CMA) to provide1-(indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine(350 mg, 32%) as an off white solid.

[1165] Melting Point=75-80° C.

[1166] IR (thin film)

[1167] 1H NMR (CDCl₃)

[1168] Analysis for C₂₈H₂₉N₄O₃: Calcd: C, 50.12; H, 5.06; N, 7.54;Found: C, 49.81; H, 5.33; N, 7.39.

[1169] HPLC Analysis (Method B): >99% t_(r)=12.4 min.

Example 3a

[1170]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)-ethyl]piperazineDihydrochloride.

[1171] Prepared from1-(indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazineusing Method I-D (but using dichloromethane as the initial solvent).

[1172] 1H NMR

[1173] IS-MS, m/e 468.2 (M+1)

[1174] Analysis for C₂₈H₂₉N₅O₂.1.9 HCl.2.0H₂O: Calcd: C, 58.70; H, 6.14;N, 12.22; Cl, 11.76; Found: C, 58.86; H, 5.62; N, 12.07; Cl, 11.78.

[1175] HPLC Analysis (Method A): 100% t_(r)=19.24 min.

Example 4

[1176]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine.

[1177] Prepared from 3-chloroindole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine using MethodI-C.

[1178] Melting point=100-105° C.

[1179] 1H-NMR (CDCl₃)

[1180] API-MS, m/e=502 (M+1)

[1181] Analysis for C₂₈H₂₈ClN₅O₂.1.2H₂O: Calcd: C, 64.23; H, 5.85; N,13.37; Found: C, 64.38; H, 5.74; N, 13.22.

[1182] HPLC Analysis (Method B): 97.2% t_(r)=13.8 min.

Example 4a

[1183]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazineDihydrochloride.

[1184] Prepared from1-(3-chloroindole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazineusing Method I-D (but using dichloromethane as the initial solvent).

[1185] 1H NMR

[1186] IS-MS, m/e 502.1 (M+1)

[1187] Analysis for C₂₈H₂₈ClN₅O₂.2.0 HCl.1.8H₂O: Calcd: C, 55.37; H,5.58; N, 11.53; Cl, 17.51; Found: C, 55.03; H, 5.34; N, 11.30; Cl,17.26.

[1188] HPLC Analysis (Method A): 100% t_(r)=24.55 min.

Example 5

[1189]1-(5-Chloroindole-2-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine.

[1190] Prepared from 5-chloroindole-2-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine using MethodI-C.

[1191] Melting Point=106-110° C.

[1192] IR (thin film)

[1193] 1H-NMR (CDCl₃)

[1194] API-MS, m/e=502 (M+1)

[1195] HPLC Analysis (Method B): 88.7% t_(r)=14.9 min.

Example 6

[1196]1-(Indole-2-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)-ethyl]piperazine.

[1197] Prepared from indole-2-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine using MethodI-C.

[1198] Melting point=95-100° C.

[1199] 1H-NMR (CDCl₃)

[1200] IR (thin film)

[1201] API-MS, m/e 468 (M+1)

[1202] Analysis for C₂₈H₂₉N₅O₂.1.7H₂O: Calcd: C, 67.51; H, 6.55; N,14.06; Found: C, 67.00; H, 6.10; N, 14.02.

[1203] HPLC Analysis (Method B): 96.5% t_(r)=13.5 min.

Example 7

[1204]1-(3-Methylindole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine.

[1205] Prepared from 3-methylindole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine using MethodI-C.

[1206] Melting point=62-65° C.

[1207] 1H-NMR (CDCl₃)

[1208] IR (thin film)

[1209] API-MS, m/e 482 (M+1)

[1210] Analysis for C₂₉H₃₁N₅O₂.1.6H₂O: Calcd: C, 68.24; H, 6.75; N,13.72; Found: C, 68.25; H, 6.66; N, 13.78.

[1211] HPLC Analysis (Method B): 93.6% t_(r)=13.3 min.

Example 8

[1212]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-[2-(2-pyridinyl)-ethyl]piperazine.

[1213] Prepared from 4-methoxybenzoyl chloride and1-(D-phenylglycinyl)-4-[2-(2-pyridinyl)ethyl]piperazine using MethodI-B.

[1214] Melting point=168-180° C.

[1215] [α]²⁵ _(D) −87.7 (c 1.00, methanol)

[1216] 1H-NMR (CD₃OD)

[1217] CI-MS, m/e=459 (M+1)

[1218] Analysis for C₂₇H₃₀N₄O₂.2.0HCl.0.9H₂O: Calcd: C, 59.21; H, 6.22;N, 10.23; Cl, 12.95, Found: C, 58.88; H, 6.25; N, 10.19; Cl, 13.26.

[1219] HPLC Analysis (Method B): 97.5% t_(r)=12.2 min.

Example 9

[1220]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-[2-(2-pyridinyl)ethyl]piperazine.

[1221] Prepared from 3-chloroindole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(2-pyridinyl)ethyl]piperazine using MethodI-C.

[1222] Melting point=93-96° C.

[1223] [α]²⁵ _(D) −72.4 (c 0.61, chloroform)

[1224] 1H-NMR (CDCl₃)

[1225] CI-MS, m/e=502 (M+1)

[1226] Analysis for C₂₈H₂₈N₅O₂.0.4H₂O: Calcd: C, 66.04; H, 5.07; N,13.75; Cl, 6.96; Found: C, 65.94; H, 5.61; N, 13.74; Cl, 6.91.

[1227] HPLC Analysis (Method B): 98.3% t_(r)=14.1 min.

Example 10

[1228]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(2-pyridinyl)-ethyl]piperazine.

[1229] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(2-pyridinyl)ethyl]piperazine using MethodI-C.

[1230] Melting point=73-78° C.

[1231] [α]²⁵ _(D) −90.9 (c 0.25, chloroform)

[1232] 1H-NMR (CDCl₃)

[1233] CI-MS, m/e=468 (M+1)

[1234] Analysis for C₂₈H₂₈N₅O₂.0.6H₂O: Calcd: C, 70.30; H, 6.36; N,14.64; Found: C, 70.39; H, 6.30; N, 14.62.

[1235] HPLC Analysis (Method B): 98.3% t_(r)=14.1 min.

Example 11

[1236]1-[4-Methoxybenzoyl-D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazineTrihydrochloride.

[1237] Prepared from 4-methoxybenzoic acid and1-[D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazine usingMethod I-C.

[1238] 1H-NMR

[1239] IS-MS, m/e 460.3 (M+1)

[1240] Analysis for C₂₆H₂₉N₅O₃.3.5 HCl.4H₂O: Calcd: C, 47.37; H, 6.19;N, 10.62; Found: C, 47.17; H, 5.75; N, 10.56.

[1241] HPLC Analysis (Method A): 100% t_(r)=10.48 min.

Example 12

[1242]1-[3-Chloroindole-6-carbonyl-D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazineTrihydrochloride.

[1243] Prepared from 3-chloroindole-6-carboxylic acid and1-[D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]-piperazine usingMethod I-C.

[1244] 1H-NMR

[1245] IS-MS, m/e 503.5 (M+1)

[1246] Analysis for C₂₇H₂₇N₆O₂Cl.3 HCl.5H₂O: Calcd: C, 46.16; H, 5.74;N, 11.96; Cl, 20.19; Found: C, 46.10; H, 5.59; N, 11.68; Cl, 20.29.

[1247] HPLC Analysis (Method A): 99% t_(r)=18.64 min.

Example 13

[1248]1-[3-Methylindole-6-carbonyl-D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazine.

[1249] Prepared from 3-methylindole-6-carboxylic acid and1-[D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]-piperazine usingMethod I-C.

[1250] 1H-NMR

[1251] IS-MS, m/e 483.5 (M+1)

[1252] HPLC Analysis (Method A): 99% t_(r)=16.14 min.

Example 14

[1253]1-[Indole-6-carbonyl-D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazine.

[1254] Prepared from indole-6-carboxylic acid and1-[D,L-(pyridin-2-yl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazine usingMethod I-C.

[1255] 1H-NMR

[1256] IS-MS, m/e 469.3 (M+1)

[1257] HPLC Analysis (Method A): 100% t_(r)=12.87 min.

Example 15

[1258]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(3-pyridinyl)-ethyl]piperazine.

[1259] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(3-pyridinyl)ethyl]piperazine using MethodI-C.

[1260] Melting point=82-87° C.

[1261] [α]²⁵ _(D) −116.0 (c 0.25, methanol)

[1262] IR (thin film)

[1263] 1H-NMR (CDCl₃)

[1264] API-MS, m/e=468 (M+1)

[1265] Analysis for C₂₈H₂₉N₅O₂.1.25H₂O: Calcd: C, 68.62; H, 6.48; N,14.29; Found: C, 68.49; H, 6.39; N, 14.13.

[1266] HPLC Analysis (Method B): >99% t_(r)=12.3 min.

Example 16

[1267]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(2-pyrazinyl)-ethyl]piperazine.

[1268] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(2-pyrazinyl)ethyl]piperazine using MethodI-C.

[1269] Melting Point=53-58° C.

[1270] [α]²⁵ _(D) −91.4 (c 0.23, chloroform)

[1271] 1H-NMR (CDCl₃)

[1272] IR (thin film)

[1273] API-MS, m/e 469 (M+1)

[1274] Analysis for C₂₇H₂₈N₆O₂.1.6H₂O: Calcd: C, 65.20; H, 6.32; N,16.90; Found: C, 65.49; H, 6.02; N, 16.54.

[1275] HPLC Analysis (Method B): 98.5% t_(r)=13.6 min.

Example 17

[1276]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(1-imidazolyl)-ethyl]piperazine.

[1277] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(1-imidazolyl)ethyl]piperazine using MethodI-C.

[1278] 1H-NMR

[1279] IS-MS, m/e 457.3 (M+1)

[1280] Analysis for C₂₆H₂₈N₆O₂.1.1H₂O: Calcd: C, 65.55; H, 6.39; N,17.64; Found: C, 66.01; H, 6.23; N, 17.14.

[1281] HPLC Analysis (Method A): 99% t_(r)=19.66 min.

Example 18

[1282]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[(2-(1-pyrazolyl)-ethyl]piperazine.

[1283] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(1-pyrazolyl)ethyl]piperazine using MethodI-C.

[1284] 1H-NMR

[1285] IS-MS, m/e 457.2 (M+1)

[1286] Analysis for C₂₆H₂₈N₆O₂.1.3H₂O: Calcd: C, 65.06; H, 6.43; N,17.51; Found: C, 65.39; H, 6.53; N, 16.98.

[1287] HPLC Analysis (Method A):>97% t_(r)=22.98 min.

Example 19

[1288]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-imidazolyl)-ethyl]piperazineDihydrochloride.

[1289] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(4-imidazolyl)ethyl]piperazine using MethodI-C.

[1290] 1H-NMR

[1291] IS-MS, m/e 457.3 (M+1)

[1292] Analysis for C₂₆H₂₈N₆O₂.2.1 HCl.4.0H₂O: Calcd: C, 51.60; H, 6.35;N, 13.89; Cl, 12.30; Found: C, 51.82; H, 6.04; N, 13.56; Cl, 12.12.

[1293] HPLC Analysis (Method A): 94% t_(r)=17.78 min.

Example 20

[1294]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyrazolyl)-ethyl]piperazineHydrochloride.

[1295] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(4-pyrazolyl)ethyl]piperazine using MethodI-C.

[1296] 1H-NMR

[1297] IS-MS, m/e 457.3 (M+1)

[1298] Analysis for C₂₆H₂₈N₆O₂.1.3 HCl.1.75H₂O: Calcd: C, 58.32; H,6.17; N, 15.70; Cl, 8.61; Found: C, 58.31; H, 5.72; N, 15.48; Cl, 8.37.

[1299] HPLC Analysis (Method A): >98% t_(r)=19.95 min.

Example 21

[1300]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(3-pyridazinyl)-ethyl]piperazine.

[1301] Prepared from indole-6-carboxylic acid and1-(D-phenylglycinyl)-4-[2-(3-pyridazinyl)ethyl]piperazine using MethodI-C.

[1302] Melting Point=219-222° C. with decomposition

[1303] 1H NMR (CDCl₃)

[1304] [α]²⁵ _(D)−53.9° (c 0.25, dimethyl sulfoxide)

[1305] API-MS, m/e=469 (M+1)

[1306] HPLC Analysis (Method B): >99% t_(r)=12.8 min.

Example 22

[1307]1-[4-Methoxybenzoyl-D,L-(2-methoxyphenyl)glycinyl]-4-[(2-(4-pyridinyl)ethyl]piperazine.

[1308] Prepared from 4-methoxybenzoic acid and1-[D,L-(2-meth-oxyphenyl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazineusing Method I-C.

[1309] 1H-NMR

[1310] IS-MS, m/e 489.1 (M+1)

[1311] Analysis for C₂₈H₃₂N₄O₄.0.5H₂O: Calcd: C, 67.59; H, 6.68; N,11.26; Found: C, 67.57; H, 6.49; N, 11.11.

[1312] HPLC Analysis (Method A): 97.2% t_(r)=16.02 min.

Example 23

[1313]1-[Indole-6-carbonyl-D,L-(2-methoxyphenyl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazineDihydrochloride.

[1314] Prepared from indole-6-carboxylic acid and1-[D,L-(2-methoxyphenyl)glycinyl]-4-[2-(4-pyridinyl)ethyl]piperazineusing Method I-C, followed by Method I-D.

[1315] 1H-NMR

[1316] IS-MS, m/e 498.0 (M+1)

[1317] Analysis for C₂₉H₃₁N₅O₃.2.1 HCl.2.5H₂O: Calcd: C, 56.25; H, 6.20;N, 11.31; Cl, 12.02; Found: C, 56.56; H, 5.83; N, 11.21; Cl, 12.13.

[1318] HPLC Analysis (Method A): 100% t_(r)=17.24 min.

Methods for Examples 24-25

[1319] The compounds of Examples 24 and 25 were prepared by couplingBoc-D-4-carboxamidophenylglycine to the appropriate amine with EDCI/HOAt(similar to Method C-C), deprotection with TFA/DCM (similar to MethodD-B) and coupling to 3-amino-4-chlorobenzoic acid with EDCI/HOAt(similar to Method I-C).

Example 24

[1320]1-[3-Amino-4-chlorobenzoyl-D-(4-carboxamidophenyl)glycinyl]-4-(2-phenylethyl)piperazine.

[1321] HPLC (Method C) rt 11.1 min.

[1322] LCMS M+1 521.

[1323] NMR

Example 25

[1324]1-[3-Amino-4-chlorobenzoyl-D-4-carboxamidophenylglycinyl)-4-benzylpiperazine.

[1325] HPLC (Method C) rt 11.4 min.

[1326] LCMS M+1 512.

[1327] NMR

Example 26

[1328]1-[Indole-6-carbonyl-D-(4-carboxyphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1329] By coupling of Boc-D-4-carboxymethylphenylglycine with1-(1-methylpiperidin-4-yl)piperazine using HOAt and EDCI (similar toMethod C-C), followed by TFA deprotection (similar to Method D-B),coupling to indole-6-carboxylic acid using HOAt and EDCI (similar toMethod I-C) followed by hydrolysis of the methyl ester with lithiumhydroxide.

[1330] HPLC (Method C) rt, 6.05 min

[1331] LCMS M+1 504

[1332] Nmr.

Example 27

[1333]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(4-piperidinylmethyl)-piperazineTrifluoroacetate.

Preparation of Starting Materials

[1334] 1-Boc-isonipecotic Acid.

[1335] Isonipecotic acid (15.0 g, 116 mmol) was dissolved in THF (300mL), water (150 mL) and 6 N NaOH (40 mL). Di-tert-butyl dicarbonate(26.6 g, 122 mmol) was added and the mixture stirred overnight. Themixture was diluted with water and ethyl acetate, and the layersseparated. The water layers were extracted with ethyl acetate, and theorganic layers discarded. The water layer was diluted with KHSO₄ (2 N,pH˜4) and extracted with ethyl acetate. The organic layer was washedwith brine, dried (Na₂SO₄), filtered and concentrated to provide1-Boc-isonipecotic acid (23.9 g, 90%) as a white solid.

[1336] 1H-NMR(CDCl₃)

[1337] API-MS, m/e=230 (M+1)

[1338] 1-Boc-piperidine-4-methanol.

[1339] 1-Boc-isonipecotic acid (10.0 g, 214 mmol) was dissolved in THF(400 mL) and cooled to 0° C. A solution of BH₃.THF (180 mL, 1 N in THF,180 mmol) was added slowly. The mixture stirred for 1 h at 0° C. and wasallowed to warm to room temperature for 12 h. The mixture was carefullyquenched with water and diluted with ethyl acetate. The water layer wasextracted with ethyl acetate. The organic layers were combined, washedwith brine, dried (Na₂SO₄), filtered and concentrated to provide1-Boc-piperidine-4-methanol (7.98 g, 85%) as a white solid.

[1340] 1H-NMR(CDCl₃)

[1341] API-MS, m/e=220 (M+1)

[1342] 1-Boc-piperidine-4-carboxaldehyde.

[1343] Dimethyl sulfoxide (3.5 mL, 48.7 mmol) was dissolved indichloromethane (100 mL) and was cooled to −78° C. Oxalyl chloride (3.65mL, 41.8 mmol) was added. The mixture stirred for 30 min. To thissolution was added a solution of 1-Boc-piperidine-4-methanol (7.5 g,34.8 mmol) in dichloromethane (15 mL), and the mixture stirred for 1 h.Triethylamine (9.7 mL, 69.6 mmol) was added slowly and the mixturestirred at −78° C. for 30 min and warmed to room temperature over thecourse of 1 h. The mixture was diluted with water and the layersseparated. The water layer was extracted with dichloromethane and theorganic layers combined, dried (Na₂SO₄), filtered and concentrated toprovide 1-Boc-piperidine-4-carboxaldehyde (6.75 g, 91%) as a yellow oil.

[1344] 1H-NMR(CDCl₃)

[1345] API-MS, m/e=214 (M+1)

[1346]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)piperazine.

[1347] Prepared from 1-(4-methoxybenzoyl-D-phenylglycinyl)-piperazinetrifluoroacetate and 1-Boc-piperidine-4-carboxaldehyde using Method I-A(but using sodium triacetoxyborohydride in 1,2-dichloroethane)(85%).

[1348] 1H-NMR(CDCl₃)

[1349]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(4-piperidinyl-methyl)piperazinetrifluoroacetate.

[1350] Prepared from1-(4-methoxybenzoyl-D-phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)piperazineusing Method H-A (90%).

[1351] Melting Point=70-72° C. with decomposition

[1352] IR(KBr)

[1353] 1H-NMR(CD₃OD)

[1354] API-MS, m/e=451 (M+1)

[1355] Analysis for C₂₆H₃₄N₄O₃.2.5 TFA-0.4H₂O: Calcd: C, 50.12; H, 5.06;N, 7.54; Found: C, 49.81; H, 5.33; N, 7.39.

[1356] HPLC Analysis (Method B): 97.1% RT=14.3 min.

Methods for Examples 28-29

[1357] Unless otherwise indicated, using Method I-A, the title compoundswere prepared from1-(4-methoxybenzoyl-D-phenylglycinyl)-4-(4-piperidinylmethyl)piperazinetrifluoroacetate and the indicated aldehyde or ketone.

Example 28

[1358]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-ylmethyl)piperazine.

[1359] Prepared from paraformaldehyde (56%).

[1360] IR (KBr)

[1361] 1H-NMR(CD₃OD)

[1362] CI-MS, m/e=465 (M+1)

Example 29

[1363]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-isopropyl-piperidin-4-ylmethyl)piperazineTrihydrochloride.

[1364] Prepared from acetone using Method I-A, followed by Method I-D(but using methanol in place of ether/dichloromethane) (72%).

[1365] Melting Point=172-180° C. with decomposition

[1366] IR (KBr)

[1367] 1H-NMR(CD₃OD)

[1368] CI-MS, m/e=493 (M+1)

[1369] Analysis for C₂₉H₄₀N₄O₃-3 HCl: Calcd: C, 55.85; H, 7.34; N, 8.98;Found: C, 55.63; H, 7.32; N, 8.66.

[1370] HPLC Analysis (Method B): 98.2% RT=14.4 min.

Example 30

[1371]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(2-cyclopentylethyl)-piperazine

[1372] Preparation of Starting Materials:

[1373] Cyclopentylacetaldehyde.

[1374] Prepared from 2-cyclopentylethanol using the Dess-Martinoxidation (Dess, D. B.; Martin, J. C.; J. Am. Chem. Soc., 1991, 113,7277). The aldehyde was used with trace amounts of ether and methylenechloride present due to volatility of product.

[1375] 1H NMR (CDCl₃)

[1376]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(2-cyclopentylethyl)-piperazine.

[1377] Prepared from cyclopentylacetaldehyde using Method I-A (58%)

[1378] 1H NMR (CDCl₃)

Example 30A

[1379]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(2-cyclopentylethyl)-piperazineHydrochloride Hydrate.

[1380] Method I-D

[1381] To a stirred solution of1-(4-methoxybenzyl-D-phenylglycinyl)-4-(2-cyclopentylethyl)piperazine(260 mg, 0.58 mmol) in ether (10 mL) and methylene chloride (1 mL) wasadded hydrogen chloride as a 2 N solution in ether (about 2 mL), and theresulting precipitate was filtered to give1-(4-methoxybenzoyl-D-phenylglycinyl)-4-(2-cyclopentyl-ethyl)piperazinehydrochloride as a pale yellow solid.

[1382] 1H NMR (CD₃OD)

[1383] IS-MS, m/e=450 (M+1)

[1384] Analysis for C₂₇H₃₅N₃O₃.HCl.0.5H₂O: Calcd: C, 65.51; H, 7.53; N,8.49; Found: C, 65.67; H, 7.58; N, 8.13.

[1385] HPLC Analysis (Method D): >99%, RT=15.84

[1386] Melting Point=190-192° C.

Example 31

[1387]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(3-pyrrolidinyl)-piperazineTrifluoroacetate.

[1388] Preparation of Starting Materials

[1389] (R)-(+)-1-Boc-3-pyrrolidinol.

[1390] To a stirred solution of (R)-(+)-3-pyrrolidinol (2 g, 22.96 mmol)in tetrahydrofuran (60 mL) and water (30 mL) was added di-tert-butyldicarbonate (5.27 g, 24.15 mmol) and 3 N sodium hydroxide (16 mL), andthe resulting solution was stirred for 6 h. Another portion ofdi-tert-butyl dicarbonate (0.74 g, 0.34 mmol) was added and the solutionwas stirred overnight. The reaction was diluted with water (40 mL) andextracted with ethyl acetate (2×150 mL). The combined organic extractswere washed with 2 N potassium hydrogen sulfate (200 mL), saturatedsodium bicarbonate (2×150 mL), brine (150 mL) and dried over magnesiumsulfate. Removal of solvent in vacuo gave (R)-(+)-1-Boc-3-pyrrolidinol(4.21 g, 98%) as a yellow oil.

[1391] 1H-NMR (CDCl₃)

[1392] 1-Boc-3-pyrrolidinone.

[1393] Prepared from (R)-(+)-1-Boc-3-pyrrolidinol using the Dess-Martinoxidation (Dess, D. B.; Martin, J. C.; J. Am. Chem. Soc., 1991, 113,7277) (85%).

[1394] 1H NMR (CDCl₃)

[1395]1-(4-Methoxybenzyl-D-phenylglycinyl)-4-(1-Boc-3-pyrrolidinyl)-piperazine.

[1396] Prepared from 1-(4-methoxybenzyl-D-phenylglycinyl)piperazinetrifluoroacetate and 1-Boc-3-pyrrolidinone using Method I-A (69%).

[1397] 1H NMR (CDCl₃)

[1398]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(3-pyrrolidinyl)-piperazineTrifluoroacetate.

[1399] Prepared from1-(4-methoxybenzyl-D-phenylglycinyl)-4-(1-Boc-3-pyrrolidinyl)piperazineusing Method H-A.

[1400] 1H NMR (CD₃OD)

Methods for Examples 32-33

[1401] Using Method I-A (but using sodium triacetoxyborohydride in1,2-dichloroethane), the title compounds were prepared from1-(4-methoxybenzoyl-D-phenylglycinyl)-4-(3-pyrrolidinyl)-piperazinetrifluoroacetate and the indicated aldehyde or ketone.

Example 32

[1402]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpyrrolidin-3-yl)piperazine.

[1403] Prepared from paraformaldehyde (20%).

[1404] 1H-NMR(CDCl₃)

Example 33

[1405]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-isopropyl-pyrrolidin-3-yl)piperazine.

[1406] Prepared from acetone (59%).

[1407] 1H-NMR(CDCl₃)

Methods for Examples 34-46

[1408] Unless otherwise indicated, the products of Examples 34-46 wereobtained from 1-(4-methoxybenzoyl-D-phenylglycinyl)piperazine and theindicated aldehyde or ketone using Method I-A.

Example 34

[1409]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(2-pyridylmethyl)-piperazine.

[1410] Prepared from 2-pyridinecarboxaldehyde (48%).

[1411] 1H-NMR

[1412] IS-MS, m/e 444.9 (M+1)

[1413] Analytical RPHPLC, Method A, RT=21.70 min (100%)

Example 35

[1414]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(3-pyridylmethyl)-piperazine.

[1415] Prepared from 3-pyridinecarboxaldehyde (42%).

[1416] 1H-NMR

[1417] IS-MS, m/e 444.9 (M+1)

[1418] Analytical RPHPLC, Method A, RT=17.84 min (99%)

Example 36

[1419]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(4-pyridylmethyl)-piperazine.

[1420] Prepared from 4-pyridinecarboxaldehyde (45%).

[1421] 1H-NMR

[1422] IS-MS, m/e 444.9 (M+1)

[1423] Analytical RPHPLC, Method A, RT=18.36 min (99%)

Example 37

[1424] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(3-pentyl)piperazine.

[1425] Prepared from 3-pentanone (88%).

[1426] 1H-NMR

[1427] IS-MS, m/e 424.0 (M+1)

[1428] Analytical RPHPLC, Method A, RT=23.62 min (100%)

Example 38

[1429] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-cyclopentylpiperazine.

[1430] Prepared from cyclopentanone (95%).

[1431] 1H-NMR

[1432] IS-MS, m/e 422.0 (M+1)

[1433] Analytical RPHPLC, Method A, RT=20.76 min (100%)

Example 39

[1434]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(4-methylcyclohexyl)-piperazine.

[1435] Prepared from 4-methylcyclohexanone (46%).

[1436] 1H-NMR

[1437] IS-MS, m/e 450.0 (M+1)

[1438] Analytical RPHPLC, Method A, RT=27.07 min (isomer 1), 27.74 min(isomer 2).

Example 40

[1439]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(tetrahydrothiopyran-4-yl)piperazine.

[1440] Prepared from tetrahydro-4H-thiopyran-4-one (86%).

[1441] 1H-NMR

[1442] IS-MS, m/e 453.9 (M+1)

[1443] Analytical RPHPLC, Method A, RT=22.96 min (100%)

Example 41

[1444] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(2-indanyl)piperazine.

[1445] Prepared from 2-indanone (92%).

[1446] 1H-NMR

[1447] IS-MS, m/e 469.9 (M+1)

[1448] Analytical RPHPLC, Method A, RT=26.32 min (100%)

Example 42

[1449] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-benzylpiperazine.

[1450] Prepared from benzaldehyde (87%).

[1451] 1H-NMR

[1452] IS-MS, m/e 444.0 (M+1)

[1453] Analytical RPHPLC, Method A, RT=25.78 min (96%)

Example 43

[1454]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(cyclohexyl-methyl)piperazine.

[1455] Prepared from cyclohexanecarboxaldehyde (86%).

[1456] 1H-NMR

[1457] IS-MS, m/e 450.2 (M+1)

[1458] Analytical RPHPLC, Method A, RT=28.07 min (94%)

Example 44

[1459] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(4-heptyl)piperazine.

[1460] Prepared from 4-heptanone (89%).

[1461] 1H-NMR

[1462] IS-MS, m/e 452.0 (M+1)

[1463] Analytical RPHPLC, Method A, RT=29.62 min (94%)

Example 45

[1464] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(4-pyranyl)piperazine.

[1465] Prepared from pyran-4-one (95%).

[1466] 1H-NMR

[1467] IS-MS, m/e 437.9 (M+1)

[1468] Analytical RPHPLC, Method A, RT=18.46 min (97.5%)

Example 46

[1469] 1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-cyclohexylpiperazine.

[1470] Prepared from cyclohexanone (quantitative).

[1471] 1H-NMR

[1472] IS-MS, m/e 436.0 (M+1)

[1473] Analytical RPHPLC, Method A, RT=23.43 min (100%)

Examples 47-50

[1474] Preparation of Starting Materials

[1475] 1-(Cbz-D-Phenylglycinyl)piperazine.

[1476] Prepared from 1-(Cbz-D-phenylglycinyl)-4-Boc-piperazine usingMethod H-A. The crude product was dissolved in ethyl acetate and washedwith satd aq. NaHCO₃, followed by brine, then dried with MgSO₄, filteredand concentrated in vacuo (85%).

[1477] 1H-NMR

[1478] IS-MS, m/e 354.2 (M+1)

[1479] Analysis for C₂₀H₂₃N₃O₃.0.2H₂O: Calcd: C, 67.28; H, 6.61; N,11.77; Found: C, 67.10; H, 6.46; N, 11.63.

[1480] 1-(Cbz-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1481] Prepared from (Cbz-D-phenylglycinyl)piperazine and1-methylpiperidin-4-one using Method I-A (but using sodiumtriacetoxyborohydride in 1,2-dichloroethane)(49%).

[1482] 1H-NMR

[1483] IS-MS, m/e 451.3 (M+1)

[1484] Analysis for C₂₆H₃₄N₄O₃: Calcd: C, 69.31; H, 7.61; N, 12.43;Found: C, 69.36; H, 7.71; N, 13.14.

[1485] 1-D-Phenylglycinyl-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1486] To a stirring suspension of 5% Pd/C (0.6 g) in ethanol (25 mL)under nitrogen was added a solution of1-(Cbz-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine (2.6 g,5.77 mmol) and acetic acid (1.6 mL) in ethanol (50 mL). The flask wasplaced under vacuum and the atmosphere was replaced with hydrogen(balloon). After 4 h, diatomaceous earth was added and the mixture wasfiltered through a pad of diatomaceous earth and concentrated in vacuo.The residue was dissolved in ethyl acetate and HCl gas was bubbledthrough the stirring solution to precipitate the dihydrochloride salt.The mixture was filtered and the solid was dried in vacuo to give 2.6 g(quantitative) of the title compound.

[1487] 1H-NMR

[1488] IS-MS, m/e 317.3 (M+1)

[1489] 1-Boc-D-Phenylglycinyl-4-(1-methylpiperidin-4-yl)piperazine.

[1490] Boc-D-Phg-OH (40.0 g, 159.2 mmol) and1-(1-methylpiperidin-4-yl)piperazine (32.1 g, 175.1 mmol) were slurriedin anhydrous dichloromethane (1.5 L) under N₂. The mixture was thencooled to −15° C. in an ice/MeOH bath. Triethylamine (26.6 mL, 191.0mmol) was added slowly, maintaining the temperature at −15° C., followedby slow addition of diethyl cyanophosphonate (29.0 mL, 191.0 mmol),again maintaining temp at −15° C. The reaction mixture was allowed towarm to room temperature overnight. The reaction was then quenched withthe addition of satd NaHCO₃ (500 mL), and the layers were separated. Theaqueous layer was then extracted with dichloromethane (3×1 L). Theorganic layers were combined, dried over Na₂SO₄, filtered andconcentrated in vacuo to give a crude oil. Purification using (Biotage)Flash Chromatography with 7.5% (2 M NH₃ in MeOH) in THF gave 53.6 g(81%) of the title compound.

[1491]¹H NMR (DMSO-d₆) δ 7.33 (m, 5H), 7.12 (d, J=8.1 Hz, 1H), 5.53 (d,J=8.1 Hz, 1H), 3.31 (m, 5H), 2.72 (d, J=11.3 Hz, 2H), 2.3 (m, 3H), 2.09(s, 3H), 2.03-1.86 (m, 2H), 1.76 (dt, J=9.7, 1.8 Hz, 2H), 1.56 (m, 2H),1.36 (s, 9H).

[1492] IS-MS, m/e 416.27 (M+1).

[1493] Chiral HPLC indicated no racemization had occurred.

[1494] 1-D-Phenylglycinyl-4-(1-methylpiperidin-4-yl)piperazineTrihydrochloride.

[1495] 1-Boc-D-phenylglycinyl-4-(1-methylpiperidin-4-yl)-piperazine(49.6 g, 119.1 mmol) was dissolved in anhydrous MeOH (1 L) and HCl (gas)was bubbled through the solution for 2 h 15 min, noting the formation ofa white precipitate. The solvents were removed in vacuo to give 48.3 g(95%) of the title compound as an off-white foam.

[1496]¹H NMR (DMSO-d₆) δ 12.08 (bs, 1H), 11.03 (bs, 1H), 8.92 (bs, 2H),8.79 (bs, 1H), 7.54 (m, 2H), 7.47 (m, 3H), 5.66 (s, 1H), 4.49 (m, 1H),4.26 (bd, 1H), 3.91 (bs, 2H), 3.5-2.8 (m, 9H), 2.69 (s, 3H), 2.4-1.8 (m,4H). IS-MS, m/e 316.24 (M+1).

[1497] General Procedure: Except as otherwise described, the product ofeach of Examples 47-50 was prepared from1-(D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazinedihydrochloride and the indicated acid using Method I-C (with EDCI inplace of DCC).

Example 47

[1498]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1499] Prepared from 4-methoxybenzoic acid (19%).

[1500] 1H-NMR

[1501] IS-MS, m/e 451.0 (M+1)

[1502] Analytical RPHPLC, Method A, RT=16.76 min (100%)

Example 47a

[1503]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1504] Prepared from1-(4-methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazineusing Method I-D (but using dichloromethane as the initial solvent).

[1505] 1H NMR

[1506] Analysis for C₂₈H₂₈ClN₅O₂.2.0 HCl.0.5H₂O: Calcd: C, 58.64; H,7.00; N, 10.22; Found: C, 58.92; H, 6.79; N, 10.19.

[1507] HPLC Analysis (Method A): 100% t_(r)=17.14 min.

Example 47 (Alternative Synthesis)

[1508]1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1509] 1-D-Phenylglycinyl-4-(1-methylpiperidin-4-yl)piperazinetrihydrochloride (5.0 g, 11.7 mmol) was slurried in anhydrousdichloromethane (100 mL). To the slurry was added triethyamine (6.9 mL,49.3 mmol), causing the solid to go into solution after approximately 15min. p-Anisoyl chloride (2.1 mL, 14.1 mmol) was added, and the reactionstirred for 1 h. The reaction was quenched with the addition of water(100 mL), and the layers were separated. The aqueous layer was extractedwith dichloromethane (3×100 mL). The aqueous layers were combined, driedover Na₂SO₄, filtered and concentrated. The crude material was purifiedtwice using (Biotage) Flash Chromatography, eluting with 5% (NH₃ inMeOH) in dichlormethene to give 1.4 g (26%) of the title compound.

[1510]¹H NMR (DMSO-d₆) δ 8.65 (d, J=7.7 Hz, 1H), 7.88 (d, J=8.8 Hz, 2H),7.35 (m, 5H), 6.97 (d, J=8.8 Hz, 1H), 6.40 (d, J=7.7 Hz, 1H), 3.80 (s,3H), 3.48 (m, 3H), 2.72 (d, J=11.3 Hz, 2H), 2.39 (m, 3H), 2.09 (s, 3H),2.02 (m, 2H), 1.77 (dt, J=1.8, 10.2 Hz, 2H), 1.59 (d, J=11.0 Hz, 2H),1.30 (m, 2H).

[1511] IS-MS m/e 450.26 (M+1).

[1512] [α]_(D) ²⁰=−87.62 (c=0.02, MeOH).

[1513] Analysis for C₂₆H₃₄N₄O₃—H₂O: Calcd: C, 66.64; H, 7.74; N, 11.96;Found: C, 66.79; H, 7.41; N, 11.94.

Example 48

[1514]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1515] Prepared from indole-6-carboxylic acid (65%).

[1516] 1H-NMR

[1517] IS-MS, m/e 460.2 (M+1)

[1518] Analytical RPHPLC, Method A, RT=16.68 min (100%)

Example 48a

[1519]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1520] May be prepared from1-(indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazineusing Method I-D (but using dichloromethane as the initial solvent).

Example 48 (Alternative Synthesis)

[1521]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1522] Indole-6-carboxylic acid (16.0 g, 99.3 mmol) and1-D-phenylglycinyl-4-(1-methylpiperidin-4-yl)piperazine trihydrochloride(42.3 g, 99.3 mmol) were slurried in anhydrous dichloromethane (1 L)under N₂. The mixture was then cooled to −15° C. in an ice/MeOH bath.Triethyamine (58.1 mL, 416.9 mmol) was added slowly, maintaining thetemperature at −15° C., followed by slow addition of diethylcyanophosphonate (18.1 mL, 119.1 mmol), maintaining the temperature at−15° C. The reaction mixture was allowed to warm to room temperatureovernight. The reaction was then quenched with the addition of satdNaHCO₃ (500 mL), and the layers were separated. The aqueous layer wasthen extracted with dichloromethane (3×500 mL). The organic layers werecombined, dried over Na₂SO₄, filtered and concentrated to give a crudeoil. Purification was performed using (Biotage) Flash Chromatography,eluting with 8.3% (2 M NH₃ in MeOH) in CHCl₃. The product containingfractions were combined and concentrated in vacuo to give 45.1 g (99%)of the title compound.

[1523]¹H NMR (DMSO-d₆) δ 11.35 (s, 1H), 8.65 (d, J=7.7 Hz, 1H), 7.98 (s,1H), 7.60-7.45 (m, 5H), 7.40-7.25 (m, 3H), 6.48 (t, J=2.0 Hz, 1H), 6.09(d, J=7.7 Hz, 1H), 3.5 (m, 3H), 2.72 (d, J=11.3 Hz, 2H), 2.40 (m, 2H),2.09 (s, 3H), 2.05 (m, 2H), 1.77 (dt, J=1.1, 10.2 Hz, 2H), 1.59 (d,J=11.3 Hz, 2H), 1.31 (m, 2H).

[1524]¹³C NMR (DMSO-d₆) δ 168.0, 166.4, 138.0, 135.1, 129.9, 128.4,128.2, 128.0, 127.6, 126.6, 119.4, 118.1, 111.5, 101.2, 79.1, 60.6,54.7, 53.7, 48.5, 48.3, 45.8, 45.4, 42.2, 27.7, 27.6.

[1525] IS-MS, m/e 459.26 (M+1).

[1526] [α]_(D) ²⁰=−73.08 (c=0.02, MeOH).

[1527] A portion of the free base was isolated from a chloroform—ethylacetate solvent system as crystalline material which was birefringent bymicroscopy. From DSC and TGA, the material was found to be a solvatecontaining 0.5 mol chloroform per mol of free base. The chloroformsolvate was found to have a broad endotherm about 148-158° C., followedby a sharper endotherm (peak at 194.4° C.) as the melting point of thedesolvated free base.

Example 48a (Alternative Synthesis)

[1528]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1529] To a solution of1-(indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine(14.5 g, 31.6 mmol) in anhydrous dichloromethane (300 mL) and anhydrousMeOH (150 mL) at 0° C. was added HCl in Et₂O (32.2 mL, 32.2 mmol). Afterapproximately 5 min, the solvents were removed in vacuo to give 15.1 g(96%) of the title compound.

[1530]¹H NMR (DMSO-d₆) δ 11.40 (s, 1H), 10.3 (bs, 1H), 8.68 (m, 1H),7.99 (s, 1H), 7.6-7.4 (m, 5H), 7.4-7.3 (m, 3H), 6.48 (s, 1H), 6.11 (d,J=7.3 Hz, 1H), 4.08 (bs, 1H), 3.6-1.5 (bm, 15H), 2.66 (s, 3H).

[1531] IS-MS, m/e 459.26 (M+1).

[1532] [α]_(D) ²⁰=−83.67 (c=0.01, MeOH).

[1533] Analysis for C₂₇H₃₃N₅O₂.1.1 HCl 1.7H₂O: Calcd: C, 61.03; H, 7.30;N, 13.18; Cl, 7.34; Found: C, 60.95; H, 6.91; N, 13.03; Cl, 7.00.

[1534] The product prepared by both the method of Example 48a andExample 48a (Alternative Synthesis) was found to be themono-hydrochloride salt and to be amorphous. Analysis by microscopyshowed glassy non-birefringent particles; and analysis by DSC failed toreveal a melting point, in agreement with amorphous material. Using amicrobalance flow system, the original material was cycled through avapor pressure isotherm determination, where the material deliquesced,then allowed to deydrate. Upon dehydration, there were formed crystalswhich were birefringent by microscopy; and a melting point of about 174°C. was demonstrated for the newly crystallized, hygroscopic material.

Example 48b

[1535]1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazineDifumarate.

[1536] The difumarate salt is conveniently prepared by dissolving thefree base in methanol or 95% ethanol and warming to about 50° C. (forexample at a concentration of 460 mg in 15 mL). Two molar equivalents offumaric acid (for example 232.2 mg) are then added (for example, as a0.25 M solution in methanol or as a suspension in 3 mL 95% ethanol).Following cooling and crystallization, and isolation and drying, theproduct is obtained as thin crystalline needles, with a sharp meltingpoint at about 213° C. by DSC.

Example 49

[1537]1-(3-Methylindole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1538] Prepared from 3-methylindole-6-carboxylic acid (50%).

[1539] 1H-NMR

[1540] IS-MS, m/e 474.3 (M+1)

[1541] Analytical RPHPLC, Method A, RT=22.20 min (98%)

Example 50

[1542]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine.

[1543] Prepared from 3-chloroindole-6-carboxylic acid (76%).

[1544] 1H-NMR

[1545] IS-MS, m/e 493.9 (M+1)

[1546] Analytical RPHPLC, Method A, RT=22.66 min (100%)

Example 51

[1547]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-(4-piperidinylmethyl)piperazineDihydrochloride.

[1548] Preparation of Starting Materials:

[1549] 1-(1-Boc-piperidin-4-ylmethyl)piperazine.

[1550] To a stirring solution of 1-Boc-piperidine-4-carboxaldehyde (2.4g, 11.3 mmol) in THF (60 mL) and acetonitrile (15 mL) was addedpiperazine (4.85 g, 56.3 mmol). After stirring for 5 h, sodiumtriacetoxyborohydride (2.87 g, 13.5 mmol) was added and the reaction wasallowed to stir overnight. The next morning, the solvents were removedby rotary evaporation and the residue was dissolved in ethyl acetate,washed twice with satd aq. NaHCO₃, followed by water, then dried overMgSO₄, filtered and concentrated in vacuo. The residue was thenchromatographed over silica gel, eluting with a step gradient of 2%through 15% (2 N ammonia/methanol) in dichloromethane. The productcontaining fractions were combined and concentrated in vacuo to give4.03 g (48%) of the title compound.

[1551] 1H-NMR

[1552] IS-MS, m/e 284.3 (M+1)

[1553]1-(Cbz-D-Phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)-piperazine.

[1554] Prepared from Cbz-D-phenylglycine and1-(1-Boc-piperidin-4-ylmethyl)piperazine using Method C-A. The titlecompound was purified by chromatography over silica gel, eluting with astep gradient of 1% to 3% (2 N ammonia/methanol in dichloromethane.

[1555] 1H-NMR

[1556] IS-MS, m/e 551.3 (M+1)

[1557] 1-(D-Phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)piperazine.

[1558] Prepared from1-(Cbz-D-phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)piperazine usingMethod F-A.

[1559] 1H-NMR

[1560] IS-MS, m/e 417.8 (M+1)

[1561]1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-(4-piperidinylmethyl)piperazineDihydrochloride.

[1562] Prepared from 3-chloroindole-6-carboxylic acid and1-(D-phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)piperazine usingMethods I-C, D-B, and I-D (using dichloromethane in place ofether/dichloromethane as an initial solvent).

[1563] 1H-NMR

[1564] IS-MS, m/e 494.2 (M+1)

[1565] Analysis for C₂₇H₃₂N₅O₂Cl.2.2 HCl.3.0H₂O: Calcd: C, 51.61; H,6.45; N, 11.15; Cl, 18.06; Found: C, 51.40; H, 6.12; N, 11.02; Cl,17.80.

[1566] Analytical RPHPLC, Method A, RT=20.59 min (100%)

Example 52

[1567]1-(3-Methylindole-6-carbonyl-D-phenylglycinyl)-4-(4-piperidinylmethyl)piperazineDihydrochloride.

[1568] Prepared from 3-methylindole-6-carboxylic acid and1-(D-phenylglycinyl)-4-(1-Boc-piperidin-4-ylmethyl)piperazine usingMethods I-C, D-B, and I-D (using dichloromethane in place ofether/dichloromethane as an initial solvent).

[1569] 1H-NMR

[1570] IS-MS, m/e 474.2 (M+1) Analysis for C₂₈H₃₅N₅O₂.2.3 HCl.4.0H₂O:Calcd: C, 53.42; H, 7.25; N, 11.13; Cl, 12.95; Found: C, 53.14; H, 6.71;N, 10.99; Cl, 13.12.

[1571] Analytical RPHPLC, Method A, RT=20.23 min (100%)

Example 53

[1572] 1-[4-Chlorobenzoyl-D-phenylglycinyl]-4-benzylpiperazineTrifluoroacetate.

[1573] Boc-D-phenylglycine (753 mg, 3 mmol), TBTU(2-(1H-(benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate)(963.3 mg, 3 mmol), diisopropylethylamine (894 mg, 6 mmol) and4-benzylpiperazine (525 mg, 3 mmol) were combined in DMF (10 mL) andstirred overnight. The reaction mixture was taken into dichloromethane(25 mL) washed with water (50 mL) and evaporated to dryness.

[1574] The residue was treated with TFA (5 mL) for 1 h and the excessTFA evaporated in vacuo. Triethylamine (1 mL) was added and evaporatedin vacuo. This mixture was then divided into three equal parts. One partwas then treated with a mixture of 4-chlorobenzoic acid (156.5 mg, 1mmol), HOBt (148.5 mg, 1.1 mmol) and EDCI (191 mg, 1 mmol) in DMF (3 mL)that had been stirred for 5 min. The reaction mixture was stirredovernight, diluted with water and acetonitile, and applied directly forpurification by preparative RPHPLC to give the title compound, (120 mg).

[1575] 1H-NMR

[1576] By similar methods to those described in Example 53 the followingcompounds were prepared:

Example 54

[1577] 1-[4-Chlorobenzoyl-D-phenylglycinyl]-4-(2-phenethyl) piperazineTrifluoroacetate.

[1578] 1H-NMR

[1579] MS MALDI TOF M+1=462

Example 55

[1580]1-[4-Chlorobenzoyl-D-phenylglycinyl]-4-(cyclohexylmethyl)-piperazineTrifluoroacetate.

[1581] 1H-NMR

[1582] MS MALDI TOF M+1=454

Example 56

[1583]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(thiazol-2-yl)-ethyl]piperazineHydrochloride.

[1584] Prepared from1-(D-phenylglycinyl)-4-[2-(thiazol-2-yl)-ethyl]piperazinetrihydrochloride and indole-6-carboxylic acid using methodssubstantially equivalent to Method I-C followed by Method I-D.

[1585] Melting Point=135-142° C. with decomposition.

[1586]¹H NMR (CD₃OD).

[1587] APCI-MS, m/e=474 (C₂₆H₂₇N₅O₂S+1).

[1588] HPLC Analysis (Method B): 98.8% t_(r)=14.2 min.

Example 57

[1589] (No Example 57)

Example 58a

[1590]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(3-fluoropyridin-4-yl)ethyl]piperazine.

[1591] Prepared from1-(D-phenylglycinyl)-4-[2-(3-fluoropyridin-4-yl)ethyl]piperazine andindole-6-carboxylic acid using a method substantially equivalent toMethod I-C (66%).

[1592]¹H NMR (CDCl₃).

[1593] APCI-MS, m/e=486 (C₂₈H₂₈FN₅O₂+1).

Example 58b

[1594]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(3-fluoropyridin-4-yl)ethyl]piperazineHydrochloride.

[1595] Prepared from1-(indole-6-carbonyl-D-phenylglycinyl)-4-[2-(3-fluoropyridin-4-yl)ethyl]piperazineusing a method substantially equivalent to Method I-D (89%).

[1596] [α]²⁵ _(D) −98.8° (c 0.30, methanol)

[1597] Melting Point=135-145° C. with decomposition.

[1598]¹H NMR (CD₃OD).

[1599] APCI-MS, m/e=486 (C₂₈H₂₈FN₅O₂+1).

[1600] TLC R_(f)=0.44 (7:3 CH₂Cl₂:CMA)

[1601] Analysis for C₂₈H₃₁N₃O₄.1.25 HCl.1.2H₂O: Calcd: C, 60.84; H,5.77; N, 12.67; Cl, 8.02; Found: C, 61.14; H, 5.86; N, 12.34; Cl, 7.88.

[1602] HPLC Analysis (Method B): 98.2% t_(r)=13.2 min.

Example 59a

[1603]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(2-cyanopyridin-4-yl)ethyl]piperazine.

Method I-E

[1604] 1-(D-Phenylglycinyl)-4-(2-(2-cyanopyridin-4-ylethyl)-piperazinetrihydrochloride (580 mg, 1.26 mmol), indole-6-carboxylic acid (205 mg,1.26 mmol), HOBt (171 mg, 1.26 mmol), and triethylamine (0.88 mL, 6.9mmol) were dissolved in DMF (20 mL). To this solution, DCC (390 mg, 1.89mmol) was added, and the mixture stirred at room temperature overnight.Ethyl acetate (100 mL) and heptane (20 mL) were added and solids removedby filtration. The solvents were removed under vacuum and residuere-dissolved in toluene/ethyl acetate (200 mL, 1:1) and solids removedby filtration. The filtrate was washed with water and brine, dried(Na₂SO₄), filtered, and concentrated under vacuum to provide crudeproduct (800 mg). The crude product was purified by chromatography(SiO₂, 1000:10:1-250:10:1 CH₂Cl₂:methanol:concentrated ammoniumhydroxide) to provide the sub-titled compound (443 mg, 71%).

[1605]¹H NMR (CDCl₃).

[1606] TLC=0.3 (200:10:1, CH₂Cl₂:methanol:concentrated ammoniumhydroxide)

Example 59b

[1607]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(2-cyanopyridin-4-yl)ethyl]piperazineHydrochloride.

[1608] Prepared from1-(indoyl-6-carbonyl-D-phenylglycinyl)-4-[2-(2-cyanopyridin-4-yl)ethyl]piperazineusing methods substantially equivalent to those described in Method I-D,(96%).

[1609] [α]²⁵ _(D) −92.00 (c 0.27, methanol)

[1610] Melting Point=156-169° C.

[1611] IR(ATR).

[1612]¹H NMR (CD₃OD).

[1613] APCI-MS, m/e=493 (C₂₉H₂₈N₆O₂+1).

[1614] Analysis for C₂₈H₃₁N₃O₄.1.1 HCl.0.9H₂O: Calcd: C, 63.44; H, 5.68;N, 15.31; Cl, 7.10; Found: C, 63.53; H, 5.68; N, 15.41; Cl, 6.88.

[1615] HPLC Analysis (Method B): 98.9% t_(r)=15.2 min.

Example 60

[1616]1-[4-Methoxybenzoyl-D,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1617] Prepared from1-[D,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineand p-anisoyl chloride using procedures substantially equivalent tothose described in Method I-B, using dichloromethane for aqueous dioxaneand TEA for K₂CO₃, followed by Method I-D.

[1618] 1H NMR

[1619] IS-MS, m/e 485.3 (M+1)

[1620] Analysis for C₂₇H₃₂N₅O₂Cl.2.2 HCl.2.0H₂O: Calcd: C, 53.14; H,6.31; N, 11.48; Cl, 18.59; Found: C, 53.04; H, 5.86; N, 11.36; Cl,18.13.

[1621] HPLC Analysis (Method A): 100% t_(r)=19.78 min.

Example 61

[1622]1-[4-Methoxybenzoyl-D-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1623]1-[4-Methoxybenzoyl-D,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine(8.20 g, 18.16 mmol) was divided into 2 g lots and each lot wasdissolved in a mixture of chloroform (1.0 mL), isopropanol (13 mL) andheptane (26 mL). These samples were then individually chromatographedusing preparative chiral HPLC (Chiralcel OD, 8×34 cm, eluting with 35%isopropanol/65% heptane with 0.4% DMEA for 21 min at a flow rate of 350mL/min). Analytical HPLC of the the racemic mixture (Chiralcel OD,4.6×250 mm, eluting with 35% isopropanol/65% heptane with 0.4% DMEA, 1.0mL/min, UV detection at 260 nm) revealed two peaks, baseline resolved.The fractions from the preparative HPLC runs containing the peak withthe shorter retention time were combined and concentrated in vacuo togive 3.71 g of isomer 1. The fractions containing the later runningisomer were combined and concentrated to give 3.80 g of isomer 2.Biological evaluation of the two samples revealed isomer 1 to be overten times more potent than isomer 2 and on that basis, isomer 1 wastentatively assigned as the D-isomer. Isomer 1 was then chromatographedover silica gel (Biotage Quad 12/25 System, 25 mm KP-Sil [32-63 umparticle size] columns, eluting with a gradient of 0-6% 2Nammonia/methanol in dichloromethane) and the product containingfractions were combined and concentrated. The residue was thenredissolved in dichloromethane and to this stirring solution was added 1M HCl in diethyl ether (4.33 mL, 4.33 mmol). The precipitate wasfiltered and dried in vacuo to give 2.3 g (49%) of the title compound.

[1624] 1H NMR

[1625] IS-MS, m/e 485.3 (M+1)

[1626] Analysis for C₂₆.H₃₃N₄O₃C_(1-1.3) HCl.0.5H₂O: Calcd: C, 57.68; H,6.57; N, 10.35; Cl, 15.06; Found: C, 57.42; H, 6.76; N, 10.06; Cl,14.89.

Example 62

[1627]1-[Indole-6-carbonyl-D,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

Method I-F

[1628] To a stirring solution ofD,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine(0.458 g, 1.31 mmol) in dichloromethane (5 mL) and DMF (2 mL) was addedindole-6-carboxylic acid (0.233 g; 1.44 mmol). The solution was cooledto 0° C. and DEPC (0.218 mL, 1.44 mmol) was added. After 24 h, thesolution was concentrated in vacuo and the residue was dissolved in 5%acetic acid/methanol and loaded onto an SCX column. The column waswashed with methanol, and then the compound was eluted with 50% (2 Nammonia/methanol) in dichloromethane. The product containing fractionswere combined and concentrated in vacuo. The residue was thenchromatographed over silica gel, eluting with a gradient of 0-5% (2 Nammonia/methanol) in dichloromethane. Again, the product containingfractions were combined and concentrated in vacuo to give 0.6 g ofoff-white solid. The HCl salt was then prepared using Method I-D to give290 mg (39%) of the title compound.

[1629] 1H NMR

[1630] IS-MS, m/e 494.2 (M+1)

[1631] Analysis for C₂₆H₃₃N₄O₃Cl.2.2 HCl.0.5H₂O: Calcd: C, 54.38; H,6.35; N, 9.76; Cl, 19.76; Found: C, 54.08; H, 6.12; N, 9.59; Cl, 19.44.

[1632] HPLC Analysis (Method A): 100% t_(r)=21.59 min.

Example 63

[1633]1-[Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1634]1-[Indole-6-carbonyl-D,L-(2-chlorophenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine(18.8 mmol theoretical) was divided into 0.5 g lots and each lot wasdissolved in a mixture of isopropanol (5 mL) and heptane (20 mL). Thesesamples were then individually chromatographed using preparative chiralHPLC (Chiralcel OD, 8×34 cm, eluting with 30% iso-propanol/70% heptanewith 0.2% DMEA for 14 min at a flow rate of 370 mL/min). Analytical HPLCof the racemic mixture (Chiralcel OD, 4.6×250 mm, eluting with 30%isopropanol/70% heptane with 0.2% DMEA, 1.0 mL/min, UV detection at 260nm) revealed two peaks, baseline resolved. The fractions from thepreparative HPLC runs containing the peak with the shorter retentiontime were combined and concentrated in vacuo to give 2.8 g of isomer 1.The fractions containing the later running isomer were combined andconcentrated to give 2.80 g of isomer 2. Biological evaluation of thetwo samples revealed isomer 1 to be about 100 times more potent thanisomer 2; and, on that basis, isomer 1 was tentatively assigned as theD-isomer. Isomer 1 (2.6 g) was then redissolved in dichloromethane, andto this stirring solution was added 1 M HCl in diethyl ether (5.26 mL,5.26 mmol). The precipitate was filtered and dried in vacuo to give 2.4g (48%) of the title compound.

[1635] 1H NMR

[1636] IS-MS, m/e 494.0 (M+1)

[1637] Analysis for C₂₇H₃₂N₅O₂Cl.2.1 HCl.0.7H₂O: Calcd: C, 59.31; H,6.36; N, 12.81; Cl, 13.62; Found: C, 59.57; H, 6.41; N, 12.42; Cl,13.31.

Example 64

[1638]1-[Indole-6-carbonyl-D,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1639] Prepared from indole-6-carboxylic acid andD,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine usingprocedures substantially equivalent to those described in Method I-Cfollowed by Method I-D. The final product was purified by preparativeRPHPLC (Vydac C18, 90% A through 65% A; A=0.01% aq. HCl,B=acetonitrile).

[1640] 1H NMR

[1641] IS-MS, m/e 511.1 (M+1)

[1642] Analysis for C₃₀H₃₄N₆O₂.1.9 HCl.3.0H₂O: Calcd: C, 56.84; H, 6.66;N, 13.26; Cl, 10.63; Found: C, 56.79; H, 6.81; N, 13.12; Cl, 10.62.

[1643] HPLC Analysis (Method A): 98.6% t_(r)=17.84 min.

Example 65

[1644]1-[3-Chloroindole-6-carbonyl-D,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1645] Prepared from 3-chloroindole-6-carboxylic acid andD,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazine usingprocedures substantially equivalent to those described in Method I-Cfollowed by Method I-D.

[1646] 1H NMR

[1647] IS-MS, m/e 545.0 (M+1)

[1648] Analysis for C₃₀H₃₃N₆O₂Cl.1.6 HCl.1.5H₂O: Calcd: C, 57.15; H,6.01; N, 13.33; Cl, 14.62; Found: C, 56.86; H, 5.64; N, 13.02; Cl,14.37.

[1649] HPLC Analysis (Method A): 100% t_(r)=25.08 min.

Example 66

[1650]1-[3-Methylindole-6-carbonyl-D,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineTrihydrochloride.

[1651] Prepared from 3-methylindole-6-carboxylic acid andD,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)-piperazine usingprocedures substantially equivalent to those described in Method I-Cfollowed by Method I-D.

[1652] 1H NMR

[1653] IS-MS, m/e 525.1 (M+1)

[1654] Analysis for C₃₁H₃₆N₆O₂.3.0 HCl-2.6H₂O: Calcd: C, 54.68; H, 6.54;N, 12.34; Cl, 15.62; Found: C, 54.41; H, 6.25; N, 12.00; Cl, 15.99.

[1655] HPLC Analysis (Method A): 99% t_(r)=23.06 min.

Example 67

[1656]1-[4-Methoxybenzoyl-D,L-(2-trifluoromethylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1657] Prepared from using procedures substantially equivalent to thosedescribed in Method I-C followed by I-D.

[1658] 1H NMR

[1659] IS-MS, m/e 519.0 (M+1)

[1660] HPLC Analysis (Method A): 98% t_(r)=20.18 min.

Example 68

[1661]1-[4-Methoxybenzoyl-D-(2-trifluoromethylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1662]1-[4-Methoxybenzoyl-D,L-(2-trifluoromethylphenyl)-glycinyl]-4-(1-methylpiperidin-4-yl)piperazine(6.14 g, 11.8 mmol) was divided into 0.5 g lots and each lot wasdissolved in a mixture of isopropanol (20 mL) and heptane (30 mL). Thesesamples were then individually chromatographed using preparative chiralHPLC (Chiralpak AD, 8×40 cm, eluting with 45% isopropanol/55% heptanewith 0.2% DMEA for 18 min at a flow rate of 450 mL/min). Analytical HPLCof the racemic mixture (Chiralpak AD, 4.6×250 mm, eluting with 45%isopropanol/55% heptane with 0.2% DMEA, 1.0 mL/min, UV detection at 260nm) revealed two peaks, baseline resolved. The fractions from thepreparative HPLC runs containing the peak with the shorter retentiontime were combined and concentrated in vacuo to give 2.9 g of isomer 1.The fractions containing the later running isomer were combined andconcentrated to give 2.8 g of isomer 2. Biological evaluation of the twosamples revealed isomer 1 to be over 100 times more potent than isomer2; and, on that basis, isomer 1 was tentatively assigned as theD-isomer. Isomer 1 (2.7 g) was then redissolved in dichloromethane andto this stirring solution was added 1 M HCl in diethyl ether (10.4 mL,10.4 mmol). The precipitate was filtered and dried in vacuo to give 2.8g (75%) of the title compound.

[1663] 1H NMR

[1664] IS-MS, m/e 520.1 (M+1)

[1665] Analysis for C₂₇H₃₃N₄O₃F.2.1 HCl.1.4H₂O: Calcd: C, 52.43; H,6.14; N, 9.06; Cl, 12.04; Found: C, 52.06; H, 6.03; N, 9.41; Cl, 11.91.

Example 69

[1666]1-[Indole-6-carbonyl-D,L-(2-trifluoromethylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1667] Prepared from1-[D,L-(2-trifluoromethylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineand 6-carboxyindole using procedures substantially equivalent to thosedescribed in Method I-C followed by I-D.

[1668] 1H NMR

[1669] IS-MS, m/e 528.0 (M+1)

[1670] Analysis for C₂₈H₃₂N₅O₂F₃1.9 HCl.2.5H₂O: Calcd: C, 52.39; H,6.10; N, 10.91; Cl, 10.50; Found: C, 52.12; H, 5.61; N, 10.71; Cl,10.63.

[1671] HPLC Analysis (Method A): 97% t_(r)=21.10 min.

Example 70

[1672]1-[4-Methoxybenzoyl-D-cyclopentylglycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

Method I-G

[1673] To a stirring solution of HOBt (0.388 g, 2.87 mmol) in DMF (2 mL)was added DCC (0.684 g, 2.65 mmol). To this solution was added asolution of p-methoxybenzoic acid (0.404 g, 2.65 mmol) and1-(D-cyclopentylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine (0.683 g,2.21 mmol) in DMF (9 mL). After stirring overnight, the solution wasfiltered; and the filtrate was loaded onto an SCX column (pretreatedwith 5% acetic acid/methanol and washed with methanol). The column waswashed with methanol and then the product was eluted with 1 Nammonia/methanol followed by dichloromethane. The product containingfractions were combined and concentrated in vacuo to give 0.687 g, (70%,¹H NMR; IS-MS, m/e 443.4 (M+1)) of the free base of the title compound.

[1674] The HCl salt was prepared using Method I-D to give 737 mg (95%)of the title compound.

[1675] 1H NMR

[1676] IS-MS, m/e 443.4 (M+1)

[1677] Analysis for C₂₅H₃₈N₄O₃.2.1 HCl.2.0H₂O: Calcd: C, 54.08; H, 8.01;N, 10.09; Cl, 13.41; Found: C, 54.35; H, 7.76; N, 10.06; Cl, 13.64.

[1678] HPLC Analysis (Method A): 99.4% t_(r)=17.84 min.

Example 71

[1679]1-[Indole-6-carbonyl-D-cyclopentylglycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1680] Prepared from indole-6-carboxylic acid and1-[D-cyclo-pentylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine usingprocedures substantially equivalent to those used in Method I-G, usingHOBt in place of HOAt and EDCI for DCC, and Method I-D.

[1681] 1H NMR

[1682] IS-MS, m/e 452.3 (M+1)

[1683] Analysis for C₂₆H₃₇N₅O₂.1.9 HCl.2.5H₂O: Calcd: C, 55.18; H, 7.82;N, 12.38; Cl, 11.90; Found: C, 55.46; H, 7.47; N, 12.35; Cl, 11.79.

[1684] HPLC Analysis (Method A): 96.7% t_(r)=17.76 min.

Example 72

[1685]1-[4-Methoxybenzoyl-D-cyclohexylglycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1686] Prepared from 4-methoxybenzoic acid and1-[D-cyclohexylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine usingprocedures substantially equivalent to those in Method I-G and MethodI-D.

[1687] 1H NMR

[1688] IS-MS, m/e 457.4 (M+1)

[1689] Analysis for C₂₆H₄₀N₄O₃.2.0 HCl.0.6H₂O: Calcd: C, 57.79; H, 8.06;N, 10.37; Cl, 13.12; Found: C, 57.54; H, 8.02; N, 10.19; Cl, 13.22.

[1690] HPLC Analysis (Method A): 100% t_(r)=19.35 min.

Example 73

[1691]1-[Indole-6-carbonyl-D-cyclohexylglycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1692] Prepared from indole-6-carboxylic acid and1-[(D-cyclo-hexylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine usingprocedures substantially equivalent to those in Method I-G, substitutingHOAt for HOBt and EDCI for DCC, and Method I-D.

[1693] 1H NMR

[1694] IS-MS, m/e 466.3 (M+1)

[1695] Analysis for C₂₇H₃₈N₅O₂.2.1 HCl.2.0H₂O: Calcd: C, 56.08; H, 7.86;N, 12.11; Cl, 12.88; Found: C, 56.29; H, 7.47; N, 12.11; Cl, 12.76.

[1696] HPLC Analysis (Method A): 97.8% t_(r)=20.08 min.

Example 74a

[1697]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-(2-phenethyl)-piperazine.

[1698] Prepared from 1-(D-phenylglycinyl)-4-(2-phenethyl)-piperazine andindole-6-carboxylic acid using methods substantially equivalent to thosedescribed in Method I-C (62%).

[1699]¹H NMR (CDCl₃).

[1700] APCI-MS, m/e=467 (M+1).

Example 74b

[1701] 1-[Indole-6-carbonyl-D-phenylglycinyl]-4-(2-phenethyl)-piperazineHydrochloride.

[1702] Prepared from1-(indole-6-carbonyl-D-phenylglycinyl)-4-(2-phenethyl)piperazine usingmethods substantially equivalent to those described in Method I-D (96%).

[1703] [α]²⁵ _(D)=−96.8° C. (c 0.25, methanol).

[1704] Melting Point=210-215° C. (dec.)

[1705]¹H NMR (CD₃OD).

[1706] HPLC Analysis (Method B): 98.6% t_(r)=16.5 min.

[1707] Analysis for C₂₉H₃₀N₄O₂.1.0 HCl.0.5H₂O: Calcd: C, 68.02; H, 6.30;N, 10.94; Cl, 6.92 Found: C, 68.21; H, 6.32; N, 10.78; Cl, 6.78

[1708] APCI-MS, m/e=467 (M+1)

Example 75

[1709]1-[4-Methoxybenzoyl-D,L-thiazol-2-ylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1710] Prepared from1-[D,L-(thiazol-2-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazinetrihydrochloride and anisoyl chloride using method I-B, substitutingN,N-di-isopropylethylamine for potassium carbonate and substitutingdichloromethane for dioxane.

[1711] 1H-NMR

[1712] LCMS m/z 458.4 (M+1)

Example 76a

[1713]1-[4-Methoxybenzoyl-D,L-(benzo[b]thiophen-3-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1714] Prepared from1-[D,L-(benzo[b]thiophen-3-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazinetrihydrochloride and anisoyl chloride using method G-A, substitutingtriethylamine for potassium carbonate and substituting dichloromethanefor dioxane.

[1715] 1H-NMR

[1716] LCMS m/z 507.4 (M+1)

Example 76b

[1717]1-[4-Methoxybenzoyl-D,L-benzothiophene-3-ylglycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1718] Prepared from1-[4-methoxybenzoyl-D,L-(benzo[b]thiophen-3-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineand 0.2 N HCl, followed by lyophilization.

[1719] LCMS m/z 507.4

Example 77a

[1720]1-[4-Methoxybenzoyl-D,L-naphthalene-1-ylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1721] Prepared from1-[D,L-(naphthalen-1-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazinetrihydrochloride and anisoyl chloride using method G-A, substitutingtriethylamine for potassium carbonate and substituting dichloromethanefor dioxane.

[1722] 1H-NMR

[1723] IS-MS m/e 501.0 (M+1)

Example 77b

[1724]1-[4-Methoxybenzoyl-D,L-naphthalene-1-ylglycinyl]-4-(1-methylpiperidin-4-yl)piperazineDihydrochloride.

[1725] Prepared from1-[D,L-(naphthalen-1-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineand 0.2 N HCl, followed by lyophilization.

[1726] 1H-NMR

[1727] LCMS m/z 501.4 (M+1)

Example 78

[1728]1-[Indole-6-carbonyl-D,L-naphthalene-1-ylglycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1729] Prepared from1-[D,L-(naphthalen-1-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazinetrihydrochloride and 6-carboxyindole using method I-C, substitutuingEDCI for DCC and substituting N,N-diisopropylamine for triethylamine.IS-MS m/e 510.0 (M+1)

Example 79

[1730]1-[4-Methoxybenzoyl-D,L-(2-methylsulfonylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineHydrochloride.

[1731] Prepared from1-[D,L-(2-methylsulfonylphenyl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazineand anisoyl chloride using Method I-B, substituting triethylamine forpotassium carbonate and substituting dichloromethane for dioxane. TheHCl salt was prepared by Method I-D, substituting ethyl acetate fordichloromethane.

[1732] 1H-NMR

[1733] IS-MS, m/e 529 (M+1)

Example 80

[1734]1-[4-Methoxybenzoyl-D,L-(thiazol-5-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazineDihydrochloride.

[1735] To a stirred solution of 4-methoxybenzoic acid (760 mg, 5.0mmol),1-[D,L-(thiazol-5-yl)glycinyl]-4-[2-(pyridin-4-yl)-ethyl]piperazine(circa 5.0 mmol) and HOAt (750 mg, 5.5 mmol) in DMF (40 mL) was addedEDCI (1.05 g, 5.5 mmol). The mixture was stirred at room temperature for20 h, and the solvent removed in vacuo. The residues were taken up inchloroform: isopropyl alcohol (2:1) and washed with satd sodiumbicarbonate. The aqueous phase was back extracted withchloroform:isopropyl alcohol (2:1) (x3), and the combined organicextracts were dried (MgSO₄) and concentrated in vacuo. The crude productwas purified by preparative RPHPLC; and the product fractionsconcentrated, taken up in chloroform:isopropyl alcohol (2:1), washedwith satd sodium bicarbonate, dried (MgSO₄) and concentrated in vacuo.The free base thus obtained was dissolved in methanol and treated with 2equivalents of HCl in ether and evaporated to dryness. The residue wasdissolved in water/acetonitrile and freeze dried to yield 786 mg of thetitle compound.

[1736] LCMS M+1 466

[1737] NMR

Example 81

[1738]1-[4-Methoxybenzoyl-D,L-(2-methylthiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazineDihydrochloride.

[1739] Prepared from1-[D,L-(2-methylthiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazineand 4-methoxybenzoic acid using methods substantially equivalent tothose described in Example 80.

[1740] LCMS M+1 480

[1741] NMR

Example 82

[1742]1-[4-Methoxybenzoyl-D,L-(2-aminothiazol-4-yl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazineDihydrochloride.

[1743] Prepared from1-(Boc-D,L-2-aminothiazol-4-ylglycinyl)-4-[2-(pyridin-4-yl)ethyl]piperazineand 4-methoxybenzoic acid using methods substantially equivalent tothose described in Example 80.

[1744] LCMS M+1 481

[1745] NNR

[1746] The following compounds are prepared using similar procedures tothose described above and the appropriate starting materials:

[1747]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(2-aminothiazol-4-yl)ethyl]piperazine.(For example by coupling indole-6-carboxylic acid with IntermediateA-10, followed by deprotection of the amino group.)

[1748]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(2-methylpyridin-4-yl)ethyl]piperazine.

[1749]1-[Indole-6-carbonyl-D-phenylglycinyl]-4-[2-(2-trifluoro-methylpyridin-6-yl)ethyl]piperazine.

[1750]1-[Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl]-4-[2-(pyridin-4-yl)ethyl]piperazine.

[1751]1-[Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl]-4-[2-(pyridazin-3-yl)ethyl]piperazine.

[1752]1-[Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl]-4-[2-(imidazol-1-yl)ethyl]piperazine.

[1753]1-[Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl]-4-[2-(imidazol-4-yl)ethyl]piperazine.

[1754]1-(Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl]-4-[2-(pyrazol-4-yl)ethyl]piperazine.

[1755]1-[4-Methoxybenzoyl-D,L-(quinolin-8-yl)glycinyl]-4-(1-methylpiperidin-4-yl)piperazine.

[1756] Assay Protocols

[1757] Enzyme Inhibition Assays:

[1758] The ability of a test compound to inhibit factor Xa may beevaluated in one or more of the following Enzyme Inhibition assays, orin other standard assays known to those skilled in the art.

[1759] Enzyme Inhibition Assay 1

[1760] Enzyme assays were carried out at room temperature in 0.1Mphosphate buffer, pH7.4 according to the method of Tapparelli et al (J.Biol. Chem. 1993,268,4734-4741). Purified human factor Xa, trypsin,thrombin and plasmin were purchased from Alexis Corporation, Nottingham,UK. Urokinase was purchased from Calbiochem, Nottingham, UK. Chromogenicsubstrates for these enzymes; pefachrome-FXA, pefachrome-TRY,pefachrome-TH, pefachrome-PL and pefachrome-UK were purchased fromPentapharm AG, Basel, Switzerland. Product (p-nitroaniline) wasquantified by adsorption at 405 nm in 96 well microplates using aDynatech MR5000 reader (Dynex Ltd, Billingshurst, UK). Km and Ki werecalculated using SAS PROC NLIN (SAS Institute, Cary, N.C., USA, Release6.11) K_(m) values were determined as 100.9 μM for factorXa/pefachrome-FXA and 81.6 μM for trypsin/pefachrome-TRY. Inhibitorstock solutions were prepared at 40 mM in Me2SO and tested at 500 μM, 50μM and 5 μM. Accuracy of Ki measurements was confirmed by comparisonwith Ki values of known inhibitors of factor Xa and trypsin.

[1761] In agreement with published data, benzamidine inhibited factorXa, trypsin, thrombin, plasmin and urokinase with Ki values of 155 μM,21 μM, 330 nM, 200 nM and 100 nM respectively. NAPAP inhibited thrombinwith a Ki value of 3 nM. Compounds of the invention were found to haveactivity in these assays.

[1762] Enzyme Inhibition Assay 2

[1763] Human factor Xa and human thrombin were purchased from EnzymeResearch Laboratories (South Bend, Ind., USA). Other proteases were fromother commercial sources. Chromogenic para-nitroanilide peptide proteasesubstrates were purchased from Midwest Biotech (Fishers, Ind., USA).

[1764] The binding affinities for human factor Xa were measured asapparent association constants (Kass) derived from protease inhibitionkinetics as described previously.^(a,b,c,d) The apparent Kass valueswere obtained using automated (BioMek-1000) dilutions of inhibitors(Kass determinations are performed in triplicate at each of four-eightinhibitor concentrations) into 96-well plates and chromogenic substratehydrolysis rates determined at 405 nm using a Thermomax plate readerfrom Molecular Devices (San Francisco). For factor Xa inhibition, theassay protocol was: 50 μl buffer (0.06 M tris, 0.3 M NaCl, pH 7.4); 25μl inhibitor test solution (in MeOH); 25 μl human factor Xa (32 nM in0.03 M tris, 0.15 M NaCl, 1 mg/ml HSA); finally, 150 μlBzIleGluGlyArgpNA (0.3 mM in water) added within 2 min to starthydrolysis. Final factor Xa was 3.2 nM. Free [Xa] and bound [Xa] weredetermined from linear standard curves on the same plate by use ofSoftmaxPro software for each inhibitor concentration and apparent Kasscalculated for each inhibitor concentration which produced hydrolysisinhibition between 20% and 80% of the control (3.2 nM factor Xa):apparent Kass=[E:I]/[E_(f)][I_(f)]=[E_(b)]/[E_(f)][I^(o)−I_(b)]. Theapparent Kass values so obtained are approximately the inverse of the Kifor the respective inhibitors [1/appKass=app Ki]. The variability ofmean apparent Kass values determined at the single substrateconcentration was +/−15%. The assay system Km was measured as0.347+/−0.031 mM [n=4]; and Vmax was 13.11+/−0.76 μm/min.

[1765] Kass values were determined with thrombin and other proteasesusing the same protocol with the following enzyme and substrateconcentrations: thrombin 5.9 nM with 0.2 mM BzPheValArgpNA; XIa 1.2 nMwith 0.4 mM pyroGluProArgpNA; XIIa 10 nM with 0.2 mM HDProPheArgpNA;plasmin 3.4 nM with 0.5 mM HDValLeuLyspNA; nt-PA 1.2 nM with 0.8 mMHDIleProArgpNA; and urokinase 0.4 nM with 0.4 mM pyroGluGlyArgpNA; aPC 3nM with 0.174 mM pyroGluProArgpNA; plasma kallikrein 1.9 nM withD-ProPheArgpNA; bovine trypsin 1.4 nM with 0.18 mM BzPheValArgpNA.

[1766] Citations

[1767] (a) Sall D J, J A Bastian, S L Briggs, J A Buben, N Y Chirgadze,D K Clawson, M L Denny, D D Giera, D S Gifford-Moore, R W Harper, K LHauser, V J Klimkowski, T J Kohn, H-S Lin, J R McCowan, A D Palkowitz, GF Smith, M E Richett, K Takeuchi, K J Thrasher, J M Tinsley, B GUtterback, S-C B Yan, M Zhang. Dibasic Benzo[b]thiophenes Derivatives asa Novel Class of Active Site Directed Thrombin Inhibitors. 1.Determination of the Serine Protease Selectivity, Structure-ActivityRelationships and Binding Orientation. J Med Chem 40 3489-3493 (1997).

[1768] (b) Smith G F, T J Craft, D S Gifford-Moore, W J Coffman, K DKurz, E Roberts, R T Shuman, G E Sandusky, N D Jones, N Chirgadze, and CV Jackson. A Family of Arginal Thrombin Inhibitors Related to Efegatran.Sem. Thrombos. Hemost. 22, 173-183 (1996).

[1769] (c) Smith G F, D S Gifford-Moore, T J Craft, N Chirgadze, K JRuterbories, T D Lindstrom, J H Satterwhite. Efegatran: A NewCardiovascular Anticoagulant. In New Anticoagulants for theCardiovascular Patient. Ed. R Pifarre. Hanley & Belfus, Inc.,Philadelphia (1997) pp 265-300.

[1770] (d) Sall D J, J A Bastian, N Y Chirgadze, M L Denny, M J Fisher,D S Gifford-Moore, R W Harper, V J Klimkowski, T J Kohn, H S Lin, J RMcCowan, M E Richett, G F Smith, K Takeuchi, J E Toth, M Zhang. DiaminoBenzo[b]thiophene Derivatives as a Novel Class of Active Site DirectedThrombin Inhibitors: 5. Potency, Efficacy and Pharmacokinetic Propertiesof Modified C-3 Side Chain Derivatives. In press, J Med Chem (1999).

[1771] In general, the compounds of formula (I) exemplified herein havebeen found to exhibit a Ki of 10 μM or less in Assay 1 and/or a Kass ofat least 0.1×10⁶ L/mole in Assay 2.

[1772] The ability of a test compound to elongate Partial ThromboplastinTime (Prothrombin Time) may be evaluated in the following testprotocols.

[1773] Partial Thromboplastin Time (Prothrombin) Test Protocol

[1774] Venous blood was collected into 3.2% (0.109 m) trisodium citratevacutainer tubes at 1 volume of anticoagulant to nine volumes of blood.The blood cells were separated by centrifugation at 700 g for tenminutes to yield plasma, which was frozen at 70° C. until required.

[1775] To perform the test, 100 μl of plasma was pipetted into in aglass test tube, 1 μl of test compound in DMSO was added, and allowed towarm to 37° over two minutes. 100 μl of warm (37°) Manchester (tissuethromboplasin) reagent (Helena Biosciences, UK) was added, allowed toequilibrate for two minutes. 100 μl of warm (37°) 25 mM calcium chloridesolution was added to initiate clotting. The test tube was tilted threetimes through a 90° angle every five seconds to mix the reagents and thetime to clot formation recorded. Data from a series of observations andtest compound concentrations are analysed by a SAS statistical analysisprogram and a CT2 (Concentration required to double clotting time) foreach compound is generated.

[1776] Compounds of the invention were found to significantly elongatethe partial thromboplastin time (Prothrombin time).

[1777] Alternative Prothrombin Time and APTT Protocols

[1778] Coagulation Determinations. Prothrombin Times and APTT valueswere determined in HUMAN PLASMA with a STA instrument (Stago). BioPT isa special non-plasma clotting assay triggered with human tissue factor(Innovin). Possible binding to albumen or to lipid was assessed bycomparing the BioPT effects in the presence/absence of 30 mg/ml humanalbumen (HSA) and 1 mg/ml phosphatidyl choline (PC). Inhibitors weredelivered in 50% MeOH vehicle.

[1779] APTT Assay

[1780] 75 μl plasma Citrol Baxter-Dade Citrated Normal

[1781] Human Plasma

[1782] 25 μl test sol'n

[1783] 75 μl Actin Baxter-Dade Activated Cephaloplastin incubate 2 minmin. @ 37°

[1784] 75 μl CaCl₂ (0.02 M)

[1785] PT Assay

[1786] 75 μl plasma

[1787] 25 μl test sol'n

[1788] 75 μl saline incubate 1 min. @ 37° C.

[1789] 75 μl Innovin Baxter-Dade Recombinant Human Tissue Factor

[1790] Compounds of the invention were found to be potent inhibitors offactor Xa.

1. A serine protease inhibitor compound of formula (I)

wherein: R₂ is a 5 or 6 membered aromatic carbon ring optionallyinterrupted by a nitrogen, oxygen or sulphur ring atom, optionally beingsubstituted in the 3 and/or 4 position (in relation to the point ofattachment of X—X) by halo, nitro, thiol, haloalkoxy, hydrazido,alkylhydrazido, amino, cyano, haloalkyl, alkylthio, alkenyl, alkynyl,acylamino, tri or difluoromethoxy, carboxy, acyloxy, MeSO₂— or R₁, orthe substituents at the 3 and 4 positions taken together form a fusedring which is a 5 or 6 membered carbocyclic or heterocyclic ringoptionally substituted by halo, haloalkoxy, haloalkyl, cyano, nitro,amino, hydrazido, alkylthio, alkenyl, alkynyl or R_(1j), and optionallysubstituted in the position alpha to the X—X group (i.e. 6 position fora six membered aromatic ring etc) by amino, hydroxy, halo, alkyl,carboxy, alkoxycarbonyl, cyano, amido, aminoalkyl, alkoxy or alkylthiowith the proviso that R₂ cannot be aminoisoquinolyl; each Xindependently is a C, N, O or S atom or a CO, CR_(1a), C(R_(1a))₂ orNR_(1a) group, at least one X being C, CO, CR_(1a) or C(R_(1a))₂; eachR_(1a) independently represents hydrogen or hydroxyl, alkoxy, alkyl,aminoalkyl, hydroxyalkyl alkoxyalkyl, alkoxycarbonyl,alkylaminocarbonyl, alkoxycarbonylamino, acyloxymethoxycarbonyl oralkylamino optionally substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl; R₁ is as defined for R_(1a), provided that R₁ is notunsubstituted aminoalkyl; Y (the α-atom) is a nitrogen atom or a CR_(1b)group; Cy is a saturated or unsaturated, mono or poly cyclic, homo orheterocyclic group, optionally substituted by groups R_(3a) orR_(3i)X_(i); each R_(3a) independently is R_(1c), amino, halo, cyano,nitro, thiol, alkylthio, alkylsulphonyl, alkylsulphenyl, triazolyl,imidazolyl, tetrazolyl, hydrazido, alkylimidazolyl, thiazolyl,alkylthiazolyl, alkyloxazolyl, oxazolyl, alkylsulphonamido,alkylaminosulphonyl, aminosulphonyl, haloalkoxy, haloalkyl, a group ofthe formula —C(X³)N(R¹¹)R¹² (wherein X³ is O or S; and R¹¹ and R¹² areindependently selected from hydrogen, methyl or ethyl or together withthe nitrogen atom to which they are attached form a pyrrolidin-1-yl,piperidin-1-yl or morpholino group), or —OCH₂O— which is bonded to twoadjacent ring atoms in Cy; X_(i) is a bond, O, NH or CH₂; R_(3i) isphenyl, pyridyl or pyrimidinyl optionally substituted by R_(3a); R_(1b),R_(1c) and R_(1j) are as defined for R_(1a); and -L-Lp(D)_(n) is of theformula:

in which R_(r) is —(CH₂)_(c)—R_(c), —CHR_(e)R_(f), —CH₂—CHR_(e)R_(f),—CH₂—CH₂—CHR_(e)R_(f), or R_(g) in which c is 1 or 2; R_(c) is thienyl,thiazolyl(which may bear an amino substituent), isothiazolyl, oxazolyl,isoxazolyl, pyrazolyl, imidazolyl, pyridyl (which may bear analkylsulphonyl, aminosulphonyl, alkylaminosulphonyl, alkylaminocarbonyl,amino, amido, (1-4C)alkoxycarbonyl, carboxy, acetylamino, chloro,fluoro, cyano, (1-3C)alkyl, trifluoromethyl, methoxy, ethoxy, nitro,hydroxy, alkylsulphonylamino, triazolyl or tetrazolyl substituent),pyrimidinyl, pyridazinyl, pyrazinyl or phenyl (which may bear a methyl,methylamino, dimethylamino, carboxy, dialkylaminosulphonyl,alkylsulphonyl, aminosulphonyl, alkylaminosulphonyl, alkylaminocarbonyl,amino, amido, alkoxycarbonyl, acetylamino, chloro, fluoro, cyano,methoxy, ethoxy, nitro, hydroxy, alkylsulphonylamino, triazolyl ortetrazolyl substituent); each of R_(e) and R_(f) independently ishydrogen or C₁₋₃alkyl; or CHR_(e)R_(f) is cyclopentyl (which may bear ahydroxy, amino, (1-3C)alkoxy, (1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy,methoxycarbonyl or ethoxycarbonyl substituent at the 3- or 4-position),cyclohexyl (which may bear a hydroxy, amino, (1-3C)alkoxy,(1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy, methoxycarbonyl orethoxycarbonyl substituent at the 3- or 4-position),tetrahydropyran-4-yl, tetrahydrothiopyran-4-yl, pyrrolidin-3-yl (whichmay bear a hydroxy, amino, (1-3C)alkoxy, (1-3C)hydroxyalkyl,(1-3C)alkyl, carboxy, methoxycarbonyl or ethoxycarbonyl substituent atthe 1-position), piperidin-4-yl (which may bear a hydroxy, amino,(1-3C)alkoxy, (1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy, methoxycarbonylor ethoxycarbonyl substituent at the 1-position), or indan-2-yl; andR_(g) is 2-methylsulphonylphenyl which may bear a 4-fluoro substituentor R_(g) is λ⁶-1,1-dioxobenzo[b]thiophen-7-yl; or aphysiologically-tolerable salt thereof; provided that Lp(D)n is not ofthe formula (K):

wherein X₂ is fluoro or hydrogen.
 2. A serine protease inhibitorcompound of formula (I)

wherein: R₂ is a 5 or 6 membered aromatic carbon ring optionallyinterrupted by a nitrogen, oxygen or sulphur ring atom, optionally beingsubstituted in the 3 and/or 4 position (in relation to the point ofattachment of X—X) by halo, nitro, thiol, haloalkoxy, hydrazido,alkylhydrazido, amino, cyano, haloalkyl, alkylthio, alkenyl, alkynyl,acylamino, tri or difluoromethoxy, carboxy, acyloxy, MeSO₂— or R₁, orthe substituents at the 3 and 4 positions taken together form a fusedring which is a 5 or 6 membered carbocyclic or heterocyclic ringoptionally substituted by halo, haloalkoxy, haloalkyl, cyano, nitro,amino, hydrazido, alkylthio, alkenyl, alkynyl or R_(1j), and optionallysubstituted in the position alpha to the X—X group (i.e. 6 position fora six membered aromatic ring etc) by amino, hydroxy, halo, alkyl,carboxy, alkoxycarbonyl, cyano, amido, aminoalkyl, alkoxy or alkylthiowith the proviso that R₂ cannot be aminoisoquinolyl; each Xindependently is a C, N, O or S atom or a CO, CR_(1a), C(R_(1a))₂ orNR_(1a) group, at least one X being C, CO, CR_(1a) or C(R_(1a))₂; eachR_(1a) independently represents hydrogen or hydroxyl, alkoxy, alkyl,aminoalkyl, hydroxyalkyl alkoxyalkyl, alkoxycarbonyl,alkylaminocarbonyl, alkoxycarbonylamino, acyloxymethoxycarbonyl oralkylamino optionally substituted by hydroxy, alkylamino, alkoxy, oxo,aryl or cycloalkyl; R₁ is as defined for R_(1a), provided that R₁ is notunsubstituted aminoalkyl; Y (the α-atom) is a nitrogen atom or a CR_(1b)group; Cy is a saturated or unsaturated, mono or poly cyclic, homo orheterocyclic group, optionally substituted by groups R_(3a) or phenyloptionally substituted by R_(3a); each R_(3a) independently is R_(1c),amino, halo, cyano, nitro, thiol, alkylthio, alkylsulphonyl,alkylsulphenyl, triazolyl, imidazolyl, tetrazolyl, hydrazido, alkylimidazolyl, thiazolyl, alkyl thiazolyl, alkyl oxazolyl, oxazolyl,alkylsulphonamido, alkylaminosulphonyl, aminosulphonyl, haloalkoxy andhaloalkyl; R_(1b), R_(1c) and R_(1j) are as defined for R_(1a); and-L-Lp(D)_(n) is of the formula:

in which R_(r) is —(CH₂)_(c)—R_(c), —CHR_(e)R_(f), —CH₂—CHR_(e)R_(f), orR_(g) in which c is 1 or 2; R_(c) is pyridyl or phenyl (which phenyl maybear a fluoro, chloro, methyl, CONH₂, SO₂NH₂, methylaminosulphonyl,dimethylaminosulphonyl, methoxy or methylsulphonyl substituent); each ofR_(e) and R_(f) independently is hydrogen or C₁₋₃alkyl; or CHR_(e)R_(f)is cyclopentyl (which may bear a methyl, ethyl or hydroxymethylsubstituent at the 3- or 4-position), cyclohexyl (which may bear amethyl, ethyl or hydroxymethyl, (1-3C)alkyl, carboxy, methoxycarbonyl orethoxycarbonyl substituent at the 3- or 4-position),tetrahydropyran-4-yl, tetrahydrothiopyran-4-yl, pyrrolidin-3-yl (whichmay bear a 1-methyl substituent), or indan-2-yl; and R_(g) is2-methylsulphonylphenyl which may bear a 4-fluoro substituent or R_(g)is λ⁶-1,1-dioxobenzo[b]thiophen-7-yl; or a physiologically-tolerablesalt thereof; provided that Lp(D)n is not of the formula (K):

wherein X₂ is fluoro or hydrogen.
 3. A compound according to claim 1wherein -L-Lp(D)_(n) is of the formula:

in which R_(r) is —(CH₂)_(c)—R_(c); in which c is 2; R_(c) is thienyl,thiazolyl (which may bear an amino substituent), isothiazolyl, oxazolyl,isoxazolyl, pyrazolyl, imidazolyl, pyridyl (which may bear an amino,methoxycarbonyl, carboxy, fluoro, cyano, methyl, methylsulphonyl,aminosulphonyl, methylaminosulfonyl, dimethylaminosulfonyl, ortrifluoromethyl substituent), pyrimidinyl, pyridazinyl, pyrazinyl orphenyl (which phenyl may bear a fluoro, chloro, cyano, methyl, amino,methylsulphonyl, aminosulphonyl, methylaminosulphonyl,dimethylaminosulphonyl, methylamino, dimethylamino, carboxy,methoxycarbonyl or methoxy substituent).
 4. A compound according to anyone of claims 1 to 3 wherein Rc is thiazolyl (which may bear an aminosubstituent), pyrimidinyl, pyrazolyl, imidazolyl, pyridyl (which maybear a methylsulphonyl, aminosulphonyl, methylaminosulfonyl,dimethylaminosulfonyl, fluoro, cyano, methyl or trifluoromethylsubstituent), pyridazinyl, pyrazinyl or phenyl (which phenyl may bear afluoro, chloro, cyano, methyl, amino, methylamino, dimethylamino,carboxy, methoxycarbonyl, methylsulphonyl, aminosulphonyl,methylaminosulfonyl, dimethylaminosulfonyl, or methoxy substituent). 5.A compound according to any one of claims 1 to 4 wherein Rc is thiazolyl(which may bear an amino substituent), pyrazolyl, imidazolyl, pyridyl(which may bear a fluoro, cyano, methyl or trifluoromethyl substituent),pyridazinyl or pyrazinyl.
 6. A compound according to any one of claims 1to 5 wherein Rc is thiazol-2-yl, 2-aminothiazol-4-yl, pyrazol-1-yl,pyrazol-4-yl, pyridazin-3-yl, imidazol-1-yl, imidazol-4-yl,pyrazin-2-yl, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl, 3-fluoropyrid-4-yl,2-cyanopyrid-4-yl, 2-methylpyrid-4-yl or 2-trifluoromethylpyrid-6-yl. 7.A compound according to claim 1 wherein -Lp(D)_(n) is of the formula:

wherein; m represents 0 or 1; X⁰ represents CH or N; and when R₃ ispresent as a substituent on an aromatic ring, it is selected fromhydrogen, alkylsulphonyl, aminosulphonyl, alkylaminosulphonyl,alkylaminocarbonyl, amino, amido, alkoxycarbonyl, acetylamino, chloro,fluoro, cyano, methoxy, ethoxy, nitro, hydroxy, alkylsulphonylamino,triazolyl and tetrazolyl; and when R₃ is present as a substituent on asaturated ring, it is selected from hydrogen, hydroxy, amino,(1-3C)alkoxy, (1-3C)hydroxyalkyl, (1-3C)alkyl, carboxy, methoxycarbonyland ethoxycarbonyl.
 8. A compound according to claim 7 wherein -Lp(D)nis of the formula:

wherein R_(i) is hydrogen or (1-6C)alkyl.
 9. A compound according to anyone of claims 1 to 8 wherein R₂ is phenyl, thien-2-yl, naphthyl,indol-2-yl, indol-6-yl, benzo[b]furan-5-yl, benzo[b]thiophen-2-yl orbenzimidazol-2-yl (each of which is optionally substituted as defined inclaim 1).
 10. A compound according to any one of claims 1 to 9 whereinoptional substituents for R₂ are selected from: fluoro, chloro, bromo,iodo, nitro, thiol, difluoromethoxy, trifluoromethoxy, hydrazido,methylhydrazido, amino, cyano, trifluoromethyl, methylthio, vinyl,ethynyl, acetylamino, carboxy, acetoxy, hydroxy, methyl, ethyl, amido(CONH₂), aminomethyl, methoxy and ethoxy.
 11. A compound according toany one of claims 1 to 10 wherein R₂ is selected from one of the formula(A′) to (H′):

wherein X₄ is O or S, R₁₃ is selected from hydrogen, fluoro, [except for(C′)] chloro or methyl and R₁₄ is selected from hydrogen, methyl, ethyl,fluoro, chloro, and methoxy and R₁₅ is selected from hydrogen, methyl,fluoro, chloro and amino.
 12. A compound according to claims 1 to 11,wherein R₂ is 4-chlorophenyl, 4-methoxyphenyl, 3-amino-4-chlorophenyl,indol-2-yl, 5-chloroindol-2-yl, indol-6-yl, 3-chloroindol-6-yl or3-methylindol-6-yl.
 13. A compound according to any one of claims 1 to12 wherein —X—X— is —CONH—.
 14. A compound according to any one ofclaims 1 to 13 wherein Y is CH.
 15. A compound according to any one ofclaims 1 to 14 wherein Cy is an optionally R_(3a) substituted: phenyl,pyridyl, thienyl, thiazolyl, naphthyl, piperidinyl, furanyl, pyrrolyl,isoxazolyl, isothiazolyl, pyrazolyl, oxazolyl, imidazolyl,1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, pyrimidinyl, pyridazinyl,quinolyl, isoquinolyl, benzofuryl, benzothienyl or cycloalkyl group, ora phenyl group substituted by R_(3i)X_(i) in which X_(i) is a bond, O,NH or CH₂ and R_(3i) is phenyl, pyridyl or pyrimidyl optionallysubstituted by R_(3a).
 16. A compound according to any one of claims 1to 14 wherein Cy is an optionally R_(3a) substituted: phenyl, pyridyl,thienyl, thiazolyl, naphthyl, piperidinyl or cycloalkyl group.
 17. Acompound according to any one of claims 1 to 16 wherein R_(3a) isselected from hydrogen, hydroxyl, alkoxy, alkyl (optionally substitutedby hydroxy, alkylamino, alkoxy, oxo, aryl or cycloalkyl), aminoalkyl(optionally substituted by hydroxy, alkylamino, alkoxy, oxo, aryl orcycloalkyl), hydroxyalkyl (optionally substituted by hydroxy,alkylamino, alkoxy, oxo, aryl or cycloalkyl), alkoxyalkyl,alkoxycarbonyl, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino(optionally substituted by hydroxy, alkylamino, alkoxy, oxo, aryl orcycloalkyl), amino, halo, cyano, nitro, thiol, alkylthio,alkylsulphonyl, alkylsulphenyl, alkylsulphonamido, alkylaminosulphonyl,aminosulphonyl, haloalkoxy, haloalkyl, a group of the formula—C(X³)N(R¹¹)R¹² (wherein X³ is O or S; and R¹¹ and R¹² are independentlyselected from hydrogen, methyl or ethyl or together with the nitrogenatom to which they are attached form a pyrrolidin-1-yl, piperidin-1-ylor morpholino group) and —OCH₂O— which is bonded to two adjacent ringatoms in Cy.
 18. A compound according to any one of claims 1 to 16wherein R_(3a) is selected from hydrogen, hydroxyl, alkoxy, alkyl(optionally substituted by hydroxy, alkylamino, alkoxy, oxo, aryl orcycloalkyl), hydroxyalkyl (optionally substituted by hydroxy,alkylamino, alkoxy, oxo, aryl or cycloalkyl), alkoxyalkyl,alkoxycarbonyl, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino(optionally substituted by hydroxy, alkylamino, alkoxy, oxo, aryl orcycloalkyl), aminoalkyl (substituted by hydroxy, alkylamino, alkoxy,oxo, aryl or cycloalkyl), amino, halo, cyano, nitro, thiol, alkylthio,alkylsulphonyl, alkylsulphenyl, alkylsulphonamido, alkylaminosulphonyl,aminosulphonyl, haloalkoxy and haloalkyl.
 19. A compound according toany one of claims 1 to 16 wherein R_(3a) is selected from hydrogen,hydroxyl, methoxy, ethoxy, methyl, ethyl, methylaminomethyl,dimethylaminomethyl, hydroxymethyl, carboxy, methoxymethyl,methoxycarbonyl, ethoxycarbonyl, methylaminocarbonyl,dimethylamino-carbonyl, aminomethyl, CONH₂, CH₂CONH₂, acetylamino,methoxycarbonylamino, ethoxycarbonylamino, t-butoxycarbonylamino, amino,fluoro, chloro, bromo, cyano, nitro, thiol, methylthio, methylsulphonyl,ethylsulphonyl, methylsulphenyl, methylsulphonylamido,ethylsulphonylamido, methylaminosulphonyl, ethylaminosulphonyl,aminosulphonyl, trifluoromethoxy, trifluoromethyl, bromo, —OCH₂O— (whichis bonded to two adjacent ring atoms in Cy) and —C(X³)N(R¹¹)R¹² (whereinX³ is O or S and R¹¹ and R¹² are independently selected from hydrogen,methyl or ethyl or together with the nitrogen atom to which they areattached form a pyrrolidin-1-yl, piperidin-1-yl or morpholino group).20. A compound according to any one of claims 1 to 16 wherein R_(3a) isselected from hydrogen, hydroxyl, methoxy, ethoxy, methyl, ethyl,methylaminomethyl, dimethylaminomethyl, hydroxymethyl, carboxy,methoxymethyl, methoxycarbonyl, ethoxycarbonyl, methylaminocarbonyl,dimethylamino-carbonyl, aminomethyl, CONH₂, CH₂CONH₂, acetylamino,methoxycarbonylamino, ethoxycarbonylamino, t-butoxycarbonylamino, amino,fluoro, chloro, cyano, nitro, thiol, methylthio, methylsulphonyl,ethylsulphonyl, methylsulphenyl, methylsulphonylamido,ethylsulphonylamido, methylaminosulphonyl, ethylaminosulphonyl,aminosulphonyl, trifluoromethoxy and trifluoromethyl.
 21. A compoundaccording to any one of claims 1 to 14 wherein Cy is selected from:

wherein: X′ is selected from O, S and NMe; X″ is selected from O and S;X′″ is selected from O, S, NH and NMe; Y′ is selected from hydrogen,amino and methyl; R_(o) is selected from hydrogen, methyl, fluoro,chloro, trifluoromethyl, methoxy, methylthio, methylsulphinyl andmethylsulphonyl; R_(m) is selected from hydrogen, methyl, fluoro,chloro, trifluoromethyl, methoxy, methylthio, methylsulphinyl,methylsulphonyl, carboxy, methoxycarbonyl and a group of the formula—C(X³)N(R¹¹)R¹² (wherein X³ is O or S and R¹¹ and R¹² are independentlyselected from hydrogen, methyl or ethyl or together with the nitrogenatom to which they are attached form a pyrrolidin-1-yl, piperidin-1-ylor morpholino group); R_(p) is selected from hydrogen and fluoro; orR_(o) and R_(m) or R_(m) and R_(p) form an —OCH₂O— group; or R_(o) andR_(m) together with the ring to which they are attached form a 5 or 6membered aryl or heteroaryl ring (wherein the heteroary ring contains 1or 2 heteroatoms selected from nitrogen, oxygen and sulfur); one ofR_(o1) and R_(o2) is hydrogen and the other is R_(o);
 22. A compoundaccording to any one of claims 1 to 14 wherein Cy is selected fromphenyl, 2-chlorophenyl, 2-methoxyphenyl, 4-carbamoylphenyl, pyrid-2-yl,pyrid-4-yl, thien-2-yl, thien-3-yl, furan-2-yl, furan-3-yl,imidazol-2-yl, thiazol-2-yl, thiazol-4-yl, thiazol-5-yl andquinolin-4-yl.
 23. A compound of the formula:

or a physiologically-tolerable salt thereof, wherein Cy, R₂ and R_(c)are as defined in any one of claims 1 to
 22. 24. A compound of theformula:

or a physiologically-tolerable salt thereof, wherein Cy and R₂ are asdefined in any one of claims 1-22.
 25. A compound as claimed in any oneof claims 1 to 24, in which the alpha atom in Y is carbon and has theconformation that would result from construction from a D-α-aminoacidNH₂—CR_(1b)(Cy)-COOH where the NH₂ represents part of X—X.
 26. Acompound as claimed in claim 1, which is selected from:1-(Indole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)-ethyl]piperazine;1-(3-Chloroindole-6-carbonyl-D-phenylglycinyl)-4-[2-(4-pyridinyl)ethyl]piperazine;1-(4-Methoxybenzoyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine;1-(Indole-6-carbonyl-D-phenylglycinyl)-4-(1-methylpiperidin-4-yl)piperazine;1-(4-Methoxybenzoyl-D-(2-chlorophenyl)glycinyl)-4-(1-methylpiperidin-4-yl)piperazine;1-(Indole-6-carbonyl-D-(2-chlorophenyl)glycinyl)-4-(1-methylpiperidin-4-yl)piperazine;and1-(4-Methoxybenzoyl-D-(2-trifluoromethylphenyl)glycinyl)-4-(1-methylpiperidin-4-yl)piperazine;and physiologically-tolerable salts thereof.
 27. A pharmaceuticalcomposition, which comprises a compound as claimed in any one of claims1 to 26 together with at least one pharmaceutically acceptable carrieror excipient.
 28. A compound as claimed in any one of claims 1 to 26,for use in therapy.
 29. Use of a compound as claimed in any one ofclaims 1 to 26 for the manufacture of a medicament for the treatment ofa thrombotic disorder.
 30. A method of treatment of a human or non-humananimal body to combat a thrombotic disorder, which comprisesadministering to said body an effective amount of a compound as claimedin claim
 1. 31. A pharmaceutical composition comprising a compound asclaimed in any one of claims 1 to 26 for use to combat a thromboticdisorder.
 32. A compound of formula I as claimed in claim 1 and named inany of the Examples herein, or a physiologically-tolerable salt thereof.33. A compound of the formula

or a salt thereof.